CN1922182B - Pyrrolotriazine compounds as kinase inhibitors - Google Patents

Abstract

The present invention provides compounds of formula (I); and pharmaceutically acceptable salts thereof. The formula (I) compounds inhibit tyrosine kinase activity of growth factor receptors such as HER1, HER2 and HER4 thereby making them useful as antiproliferative agents. The formula (I) compounds are also useful for the treatment of other diseases associated with signal transduction pathways operating through growth factor receptors.

Description

Pyrrolotriazine compounds as kinase inhibitors

This application claims priority to US Provisional Application No. 60/533,335, filed December 29, 2003, the entire disclosure of which is incorporated herein by reference.

technical field

The present invention relates to compounds that inhibit the tyrosine kinase activity of growth factor receptors such as HER1, HER2 and HER4, thereby rendering them useful as anticancer agents. The compounds are also useful in the treatment of diseases other than cancer which are associated with signal transduction pathways operated by growth factor receptors such as HER1, HER2 and HER4.

Background technique

Receptor tyrosine kinases (RTKs) are important in the transmission of biochemical signals across the plasma membrane of cells. These transmembrane molecules characteristically consist of an extracellular ligand-binding domain linked by a segment in the plasma membrane to an intracellular tyrosine kinase domain.

The human epidermal growth factor receptor (HER) family consists of four distinct receptor tyrosine kinases called HER1, HER2, HER3 and HER4. These kinases are also known as erbB1, erbB2, and the like. HER1 is also commonly known as the epidermal growth factor (EGF) receptor. With the exception of HER3, these receptors have intrinsic protein kinase activity specific for tyrosine residues of phosphoacceptor proteins. HER kinases are expressed in most epithelial cells as well as in tumor cells of epithelial origin. They are also frequently expressed in neoplastic cells of mesenchymal origin such as sarcomas or rhabdomyosarcomas. RTKs such as HER1 and HER2 are involved in cell proliferation and are associated with diseases such as psoriasis and cancer. Disruption of signaling through inhibition of these kinases would have antiproliferative and therapeutic effects.

The enzymatic activity of receptor tyrosine kinases can be stimulated by overexpression or by ligand-mediated dimerization. The HER family of receptors has been demonstrated for homodimer and heterodimer formation. An example of homodimerization is HER1 (the EGF receptor) by one of the EGF family of ligands, which includes EGF, transforming growth factor alpha, betacellulin, heparin-bound EGF, and epiregulin. ) dimerization. Heterodimerization among the four HER receptor kinases can be facilitated by members of the heregulin (also known as neuregulin) family that bind to ligands. This heterodimerization, as involving HER2 and HER3, or the HER3/HER4 combination, results in a significant stimulation of the tyrosine kinase activity of the receptor dimer, even though one of the receptors (HER3) is enzymatically inert. The kinase activity of HER2 has also been shown to be activated by overexpression of the receptor alone in various cell types. Activation of receptor homodimers and heterodimers results in phosphorylation of tyrosine residues on the receptor and on other intracellular proteins. This is followed by the activation of intracellular signaling pathways such as those involving microtubule-associated protein kinases (MAP kinases) and phosphatidylinositol 3-kinases (PI3 kinases). Activation of these pathways has been shown to lead to inhibition of cell proliferation and apoptosis. Inhibition of HER kinase signaling has been shown to inhibit cell proliferation and survival.

All protein kinases contain a structurally conserved catalytic domain of approximately 250-300 amino acid residues ¹ . Figure 1 shows the X-ray structure of HER1 ² , which contains highly conserved features of all members of the protein kinase family. The protein kinase fold is divided into two subdomains, or lobes. The smaller N-terminal lobe, or N-lobe, consists of five-stranded β sheets and a prominent α-helix, the so-called helix αC. The C leaves are larger and mainly spiral. The two lobes are connected by a single polypeptide chain (connector/hinge region), which acts as a hinge by which the two domains can rotate relative to each other upon binding of ATP and/or substrate. ATP is bound in the deep cleft between the two lobes and under a highly conserved loop connecting strands β1 and β2. This phosphate-binding loop, or P-loop, contains a conserved glycine-rich motif (GXGXψG), where ψ is usually tyrosine or phenylalanine. Glycine residues allow the loop to be in very close proximity to the ATP phosphate and coordinate through backbone interactions. This conserved aromatic side chain caps the site for phosphate transfer. ATP is fixed to the enzyme by hydrogen bonding between its adenine moiety and the backbone atoms of the linker region and its ribose ring to the initial residue of the C-terminal region.

Most preferably, phosphotransfer requires a precise spatial arrangement of several catalytic residues that are absolutely conserved in all known kinases. Asp813 and Asn818 (HER1 numbering as given in reference 2 or as Asp837 and Asn842 as found in REFSEQ: accession NM_005228) from a highly conserved loop structure at the base of the active site, the so-called The catalytic ring is released. Asp813 interacts with the attacking hydroxyl side chain of the substrate, while Asn818 participates in the hydrogen-bonding interaction that orients Asp813. Asn818 and another absolutely conserved catalytic residue, Asp831 (numbered as Asp855 as found in REFSEQ: Accession NM_005228), is also required for binding between two divalent metal cations involving coordination of a triphosphate group.

Many structures bound to different protein kinases in complex with ATP, its analogs, or small molecule inhibitors have provided a clear description of the catalytic domain and the ATP-binding cleft and the similar and different organizations that exist in the binding domain3 ^. It is now clear that there are regions within the binding cleft that are not occupied by ATP, and these show structural differences between kinase family members. Figure 2 shows the interaction of ATP with the hinge region of human cyclin-dependent kinase 2 (CDK2) ⁴ . The total region of all known kinase ATP binding sites is depicted in the figure, such as: (1) adenine binding region; (2) ribose pocket (ribose pocket); (3) phosphate binding pocket (phosphate binding pocket); (4) ) Mostly hydrophobic region 1, after the adenine loop, and (5) region 2, the cleft or tunnel adjacent to the ribose pocket and the N3 nitrogen of the adenine directed toward the surface-exposed region of the kinase domain. The effective structure of the kinase/inhibitor complex (complex) suggests that regions not occupied by ATP, such as regions 1 and 2, can be exploited to increase binding interactions and thus binding potency, and possibly also to tune the Sequence-differential selectivity among kinases.

A combination of crystallographic, modeling, screening and medicinal chemistry efforts has led to an understanding of how pyrrolotriazine chemotypes bind at the ATP binding site. Based on the X-ray crystallographic structure of pyrrolotriazine chemotype inhibitors in VEGFR-2, it has been shown that the pyrrolotriazine ring binds in the adenine pocket and enables several major interactions with the hinge region similar to ATP. In this binding mode, the C5 group is directed into the highly conserved ribose-phosphate pocket. The C4 group, depending on its chemical constituency, can be directed into specificity region 1 and the C6 group into specificity region 2. Modeling of an enumerated example of this chemotype in HER1 shows that the claimed C5 group of the present invention can occupy at least the ribose-phosphate pocket and associate with at least one or more absolutely conserved residues involved in phosphate binding , for example, Asn818 and Asp831 (HER1 numbering) interact.

The conserved nature of the catalytic core structure of kinases makes them excellent targets for total kinase inhibitor templates provided by the pyrrolotriazine ring and C5 group. This template can be successfully derivatized to make specific and potent ATP-competitive inhibitors of kinases by targeting poorly conserved regions of the ATP-binding site.

It has surprisingly been found that the compounds of the present invention and others such as those disclosed in U.S. Patent Nos. 5,457,105, 5,616,582 and 5,770,599, which contain small aniline derivatives such as bicyclic C4 positions without substituents, exhibit HER1 and HER2 activity.

references

(1) S.K.Hanks and T.Hunter, Protein kinases 6. The eukaryotic protein kinase superfamily: kinase (catalytic) domain structure and classification. FASEBJ.9(1995), pp.576-596.

(2) PDB ID: 1M14

Stamos, J., Sliwkowski, M.X., Eigenbrot, C.: Structure of the Epidermal Growth Factor Receptor Kinase Domain Alone and in Complex with a4-Anilinoquinazoline Inhibitor. J. Biol. Chem. 277 pp. 46265 (2002).

(3) H.M.Berman, J.Westbrook, Z.Feng, G.Gilliland, T.N.Bhat, H.Weissig, I.N.Shindyalov, P.E.Bourne.The Protein Data Bank.Nucleic Acids Research, 28 pp.235-242(2000): website: http://www.pdb.org/.

(4) PDB ID: 1HCK

Schulze-Gahmen, U., De Bondt, H.L., Kim, S.H.: High-resolution crystal structures of human cyclin-dependent kinase 2 with and without ATP: boundwaters and naturalligand as guides for inhibitor design. J Med Chem 39pp.49640)( .

Description of drawings

Figure 1 depicts the X-ray structure of HER1, with key parts of the typical kinase being color-coded.

Figure 2 depicts the X-ray structure of CDK2 in complex with ATP. Different regions of a typical ATP-binding site are depicted.

Detailed description of the invention

The present invention provides compounds of formula I, pharmaceutical compositions using the compounds and methods of using the compounds.

According to the present invention discloses a compound of formula I or a pharmaceutically acceptable salt or stereoisomer thereof,

where the symbols have the following meanings and are independently selected for each occurrence:

^R is cycloalkyl or substituted cycloalkyl, aryl or substituted aryl, heterocyclyl or substituted heterocyclyl;

^R is aryl, substituted aryl, heteroaryl or substituted heteroaryl, heterocyclyl or substituted heterocyclyl;

R ³ is hydrogen, alkyl or substituted alkyl;

X is a direct bond, -NR ³ - or -O-;

Y is a direct bond, alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl;

with the proviso that ^R is not indazolyl or substituted indazolyl.

These compounds inhibit the tyrosine kinase activity of growth factor receptors such as HER2.

In another specific embodiment, the present invention comprises a compound of formula I or a pharmaceutically acceptable salt or stereoisomer thereof, wherein

R ¹ is a heterocyclic group or a substituted heterocyclic group;

^R is aryl, substituted aryl, heteroaryl or substituted heteroaryl;

^R3 is hydrogen;

X is -NR ³ - or -O-;

Y is an alkyl group or a substituted alkyl group.

唑基、噻吩基、吡啶基、噻唑基、吡嗪基和苯基，其全部可适当地被一个或多个取代基所取代。Preferred ^R2 substituents include

Azolyl, thienyl, pyridyl, thiazolyl, pyrazinyl and phenyl, all of which may be suitably substituted with one or more substituents.

Preferred ^R substituents include benzyl, imidazolyl-ethyl, (methyl-imidazolyl)-ethyl, piperidinyl-ethyl, pyridyl-propyl, pyridyl-methyl, morpholinyl- Ethyl, (methyl-imidazolyl)-methyl, pyridyl-ethyl, amino-piperidinyl-methyl, 4-amino-1-methyl-piperidin-3-ol, (methyl-piperidinyl (azinyl)-ethyl, pyridyl-ethyl, (methyl-piperidinyl)-ethyl, (methyl-imidazolyl)-propyl, (methyl-piperidinyl)-methyl, (methyl base-piperazinyl)-propyl, diisopropylamino-ethyl, piperidinyl-propyl, dimethylamino-ethyl, dimethylamino-propyl, [(trifluoro-acetyl) -piperidinyl]-propyl, piperidinyl-ethyl, piperazinyl-ethyl, piperazinyl-propyl, pyrrolidinyl-ethyl, triazolyl-ethyl, triazolyl-propyl , (Dimethylamino-ethoxy)-ethyl, imidazolyl-propyl, [(trifluoro-acetyl)-piperidinyl]-propyl, (piperazinyl-ethoxy)-ethyl , [(trifluoro-acetyl)-piperazinyl]-propyl, [(trifluoro-acetyl)-piperazinyl]-ethyl, piperidinyl-methyl, pyrazolyl-ethyl, ( Amino-ethoxy)-ethyl, (methoxy-ethoxy)-ethyl, pyrazolyl-propyl, [(methoxy-ethyl)-methyl-amino]-ethyl, methoxy Linyl-propyl, (cyanomethyl-piperazinyl)-ethyl, [(cyano-ethyl)-methyl-amino]-ethyl, [(methoxy-ethyl)-piperidine base]-methyl, [(methoxy-ethyl)-piperidinyl]-ethyl, [(fluoro-ethyl)-methyl-amino]-ethyl, [(fluoro-ethyl) -Methyl-amino]-propyl, (methyl-piperidinyl)-propyl, [(methylsulfonyl-ethyl)-piperazinyl]-ethyl, [(cyano-ethyl)-piperidinyl Azinyl]-ethyl, [(methoxy-ethyl)-piperazinyl]-ethyl, [(methoxy-ethyl)-methyl-amino]-propyl, (cyanomethyl- Methyl-amino)-propyl, (cyanomethyl-methyl-amino)-ethyl, [(methylsulfonyl-ethyl)-methyl-amino]-propyl, (difluoro-piperidinyl )-propyl, (difluoro-piperidinyl)-ethyl, [(cyano-ethyl)-methyl-amino]-propyl, [(methylsulfonyl-ethyl)-methyl-amino] -Ethyl, [(trifluoro-ethyl)-piperazinyl]-ethyl, [cyanomethyl-(methylsulfonyl-ethyl)-amino]-propyl, [cyanomethyl-(methyl Sulfonyl-ethyl)-amino]-ethyl, (cyanomethyl-piperazinyl)-propyl, [(methylsulfonyl-ethyl)-piperazinyl]-propyl, [(cyano- Ethyl)-piperazinyl]-propyl, [(trifluoro-ethyl)-piperazinyl]-propyl, (methylsulfonyl-ethyl-amino)-ethyl, [(cyano-ethyl )-piperidinyl]-methyl, (cyanomethyl-piperidinyl)-methyl, (hydroxy-piperidinyl)-propyl, [(methylsulfonyl-ethyl)-piperidinyl]- Methyl, piperidinyl-methyl, piperidinyl, imidazolyl-propyl, 1-methyl-[1,4]-diazepan-6-ol, methylsulfonyl-propyl, ( First Sulfonyl-ethyl-amino)-propyl, pyrrolidinyl-methyl, methylsulfonyl-ethyl, (cyanomethyl-amino)-ethyl, (cyanomethyl-amino)-propyl, (Dioxo-thiomorpholinyl (thiomorpholinyl))-propyl, (oxo-piperidinyl)-propyl, [(difluoro-ethyl)-methyl-amino]-ethyl, morpholinyl -methyl, (hydroxy-pyrrolidinyl)-propyl, (hydroxy-piperidinyl)-propyl, pyrrolidinyl-methyl, (hydroxy-pyrrolidinyl)-propyl, methyl-piperidinyl , (methyl-pyrrolidinyl)-methyl, morpholinyl-methyl, pyrrolidinyl-methyl, (methyl-tetrahydro-pyridyl)-methyl, (cyano-ethyl)-piper Pyridyl, azetidinyl, (methylsulfonyl-ethyl)-piperidinyl, (cyano-methyl)-piperidinyl, isopropyl-piperidinyl, propyl-piperidinyl, Acetyl-piperidinyl, ethyl-piperidinyl, allyl-piperidinyl, tetrahydro-pyranyl, (hydroxy-ethyl)-piperidinyl, (methyl-pyrrolidinyl)-methyl Base, (methoxyethyl)-piperidinyl, piperidinyl, (methoxy-ethyl)-azetidinyl, (methoxy-methoxymethyl-ethyl)-piperidinyl Pyridyl, (methoxy-acetyl)-piperidinyl, methoxycarbonyl-piperidinyl, (hydroxy-acetyl)-piperidinyl, piperidine-carboxylic acid-acetoxy-ethyl, piperidine Pyridine-carboxylic acid-acetoxy-methyl-ethyl, hydroxy-piperidinyl, amino-cyclohexyl, piperidinyl, piperidine-carboxylic acid-methyl-oxo-dioxolyl ( dioxolyl)methyl, hydroxymethyl-piperidinyl, (aminomethyl)-cyclohexyl, amino-methyl-cyclohexyl, hydroxy-piperidinyl-methyl, morpholinyl, amino-cyclohexyl, hydroxymethyl Base-piperidyl, tetrahydro-pyranyl, methanesulfonyl-propyl, amino-methyl-propyl, amino-cyclohexyl, amino-methyl-cyclohexyl, (hydroxy-piperidyl)-propyl Base, piperidinyl, amino-propyl, morpholinyl-methyl, piperidinyl, (tert-butoxycarbonyl-morpholinyl)-methyl, benzyl, imidazolyl-ethyl, piperidinyl -ethyl, methoxyethyl, (diethylamino)-(methoxyethyl), pyrrolidinyl-ethyl, acetamide and methyl.

In another specific embodiment, the present invention comprises a compound of formula II or a pharmaceutically acceptable salt or stereoisomer thereof,

in

X is a direct bond, -NR ³ - or -O-;

Z is or -NR ⁷ -;

^R is aryl or substituted aryl, heteroaryl or substituted heteroaryl,

R ³ , R ⁴ and R ⁵ are independently selected from hydrogen, alkyl and substituted alkyl;

R ⁶ , R ^6a and R ^6b are independently selected from one or more of hydrogen, halogen, alkyl, alkoxy, aryloxy, -CN, -NH ₂ , -OH, -COOH, -CH ₂ OR ⁵ , -CONHSO ₂ R ⁵ , -CONR ⁴ R ⁵ , -NH Alkyl, -NHCO Alkyl, -NR ⁴ SO ₂ Alkyl, -NR ⁴ SO ₂ NR ⁴ R ⁵ , -OCONR ⁴ R ⁵ , -CF ₃ and -OCF ₃ , two of which may be attached to the same ring carbon atom, provided that the resulting compound is chemically stable;

R ⁷ is hydrogen, alkyl or -NH ₂ , and

n is 0, 1, 2 or 3.

In another specific embodiment, the present invention comprises a compound of formula III or a pharmaceutically acceptable salt or stereoisomer thereof,

in

X is a direct bond, -NR ³ - or -O-;

^R is aryl or substituted aryl, heteroaryl or substituted heteroaryl,

R ³ , R ⁴ and R ⁵ are independently selected from hydrogen, alkyl and substituted alkyl;

R ⁶ , R ^6a and R ^6b are independently selected from one or more of hydrogen, halogen, alkyl, alkoxy, aryloxy, -CN, -NH ₂ , -OH, -COOH, -CH ₂ OR ⁵ , -CONHSO ₂ R ⁵ , -CONR ⁴ R ⁵ , -NH Alkyl, -NHCO Alkyl, -NR ⁴ SO ₂ Alkyl, -NR ⁴ SO ₂ NR ⁴ R ⁵ , -OCONR ⁴ R ⁵ , -CF ₃ and -OCF ₃ , two of which may be attached to the same ring carbon atom, provided that the resulting compound is chemically stable; and

n is 0, 1, 2 or 3.

In another specific embodiment, the present invention comprises a compound of formula III,

in

唑、被取代的唑、噻唑、被取代的噻唑、吡嗪基或被取代的吡嗪基； ^R is phenyl, substituted phenyl, pyridyl, substituted pyridyl, pyrimidyl, substituted pyrimidyl,

Azole, substituted oxazole, thiazole, substituted thiazole, pyrazinyl or substituted pyrazinyl;

R ⁶ , R ^6a and R ^6b are independently selected from one or more of hydrogen, -NH ₂ , OH, alkoxy, -CONR ⁴ R ⁵ , -NR ⁴ SO ₂ alkyl, -NR ⁴ SO ₂ NR ⁴ R ^5. -OCONR ⁴ R ⁵ , -NH alkyl and -NHCO alkyl;

X is -NH-; and

n is 1 or 2.

Preferred compounds of the invention include the following

5-[(4-amino-1-piperidinyl)methyl]-N-(3-chloro-4-fluorophenyl)pyrrolo[2,1-f][1,2,4]triazine- 4-amine,

5-[(4-amino-1-piperidinyl)methyl]-N-2-naphthylpyrrolo[2,1-f][1,2,4]triazin-4-amine,

5-[(4-amino-1-piperidinyl)methyl]-N-phenylpyrrolo[2,1-f][1,2,4]triazin-4-amine,

5-[(4-Amino-1-piperidinyl)methyl]-N-(3-methoxyphenyl)pyrrolo[2,1-f][1,2,4]triazine-4- amine,

5-[(4-Amino-1-piperidinyl)methyl]-N-(3-ethynylphenyl)pyrrolo[2,1-f][1,2,4]triazin-4-amine ,

5-[(4-aminopiperidin-1-yl)methyl]-N-(4-fluoro-3-methoxyphenyl)pyrrolo[2,1-f][1,2,4]tri Azin-4-amine,

(3R,4R)-4-amino-1-[[4-[(3-chloro-4-fluorophenyl)amino]pyrrolo[2,1-f][1,2,4]triazine-5 -yl] methyl] piperidin-3-ol,

(3S,4S)-4-amino-1-[[4-[(3-chloro-4-fluorophenyl)amino]pyrrolo[2,1-f][1,2,4]triazine-5 -yl] methyl] piperidin-3-ol,

(3R,4R)-4-amino-1-[[4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl ]methyl]piperidin-3-ol,

(3S,4S)-4-amino-1-[[4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl ]methyl]piperidin-3-ol,

(3R,4R)-4-amino-1-[[4-[(3-methoxy-4-fluorophenyl)amino]pyrrolo[2,1-f][1,2,4]triazine -5-yl]methyl]piperidin-3-ol,

(3R,4R)-4-amino-1-({4-[(3-ethynylphenyl)-amino]pyrrolo[2,1-f]-[1,2,4]-triazine-5 -yl}-methyl)piperidin-3-ol,

(3R,4R)-4-amino-1-({4-[(3-ethoxyphenyl)-amino]-pyrrolo[2,1-f]-[1,2,4]triazine- 5-yl}-methyl)piperidin-3-ol

(3R,4R)-4-amino-1-{[4-(2-naphthylamino)-pyrrolo[2,1-f][1,2,4]triazin-5-yl]methyl} piperidin-3-ol,

(3R,4R)-4-amino-1-({4-[(3-methoxy-4-methyl-phenyl)amino]pyrrolo[2,1-f][1,2,4] Triazin-5-yl}-methyl)piperidin-3-ol,

(3R,4R)-4-amino-1-({4-[(3-bromophenyl)amino]pyrrolo[2,1-f][1,2,4]-triazin-5-yl} Methyl)-piperidin-3-ol,

(3R,4R)-4-amino-1-({4-[(3-fluoro-5-methoxy-phenyl)amino]pyrrolo[2,1-f][1,2,4]tri Azin-5-yl}-methyl)piperidin-3-ol,

(3S, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidin-3-ol,

(3R, 4S)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidin-3-ol,

(3S, 4R)-4-amino-1-({4-[(3-chlorophenyl)amino]-pyrrolo[2,1-f][1,2,4]-triazin-5-yl }methyl)-piperidin-3-ol,

(3S, 4R)-4-amino-1-({4-[(3-chloro-4-fluoro-phenyl)amino]pyrrolo[2,1-f][1,2,4]triazine- 5-yl}-methyl)piperidin-3-ol,

(3S, 4R)-4-amino-1-({4-[(3-ethynylphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl} Methyl)piperidin-3-ol,

(3R,4S)-4-amino-1-({4-[(3-ethynylphenyl)-amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidin-3-ol,

(3R,4S)-4-amino-1-({4-[(3-chlorophenyl)amino]pyrrolo[2,1-f][1,2,4]-triazin-5-yl} Methyl)-piperidin-3-ol,

(3R, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidin-3-ylcarbamate,

(3R,4R)-4-amino-1-({4-[(3-ethynylphenyl)-amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }-methyl)piperidin-3-ylcarbamate,

(3R, 4R)-4-amino-1-({4-[(3-chloro-4-fluoro-phenyl)amino]pyrrolo[2,1-f][1,2,4]triazine- 5-yl}-methyl)piperidin-3-ylcarbamate,

(3S, 4R)-4-amino-1-({4-[(3-chloro-4-fluoro-phenyl)amino]pyrrolo[2,1-f][1,2,4]triazine- 5-yl}-methyl)piperidin-3-ylcarbamate,

(3S, 4R)-4-amino-1-({4-[(3-ethynylphenyl)-amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }-methyl)piperidin-3-ylcarbamate,

(3S, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)-3-methylpiperidin-3-ol,

(3R/S, 5R/S)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazine -5-yl}methyl)piperidine-3,5-diol,

(3S,5S)-4-amino-1-({4-[(4-fluoro-3-methoxy-phenyl)amino]pyrrolo[2,1-f][1,2,4]tri Azin-5-yl}-methyl)piperidine-3,5-diol,

(3R,5R)-4-amino-1-({4-[(3-ethynylphenyl)-amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidine-3,5-diol,

5-{[(3R,4R)-4-amino-3-methoxypiperidin-1-yl]methyl}-N-(3-methoxyphenyl)pyrrolo[2,1-f] [1,2,4]triazin-4-amine,

5-(((4aR,8aR)-rel-hexahydro-1H-pyrido (pyrido)[3,4-b][1,4] Azin-6(7H)-yl)methyl)-N-(3-methoxyphenyl)pyrrolo[1,2-f][1,2,4]triazin-4-amine,

(3R, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)-N-(methylsulfonyl)piperidine-3-carboxamide,

(3R,4R)-4-amino-1-({4-[(3-ethynylphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl} Methyl)-N-methylpiperidine-3-carboxamide,

(3R, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)-N-methylpiperidine-3-carboxamide,

(3R, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidine-3-carboxamide,

((3R, 4R)-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazin-5-yl)methyl)- 4-((R)-1-phenylethylamino)piperidin-3-yl)methanol,

N-[(3R,4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazine- 5-yl}methyl)piperidin-3-yl]urea,

N-(3R,4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazine-5 -yl}methyl)piperidin-3-yl]methanesulfonamide, and

N-{(3S,4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazine- 5-yl}methyl)piperidin-3-yl]methanesulfonamide,

or a pharmaceutically acceptable salt thereof.

The following are definitions of terms used in this specification. Unless otherwise indicated, the initial definition of a group or term provided herein applies in this specification alone or as part of another group.

The term "alkyl" refers to a straight or branched unsubstituted hydrocarbon group of 1 to 20 carbon atoms, preferably 1 to 7 carbon atoms. The term "lower alkyl" refers to an unsubstituted alkyl group of 1 to 4 carbon atoms.

The term "substituted alkyl" refers to an alkyl group substituted with, for example, one to four of the following substituents: halogen, hydroxy, alkoxy, oxo, alkanoyl, aryloxy, alkanoyloxy, amino , alkylamino, arylamino, aralkylamino, disubstituted amines, wherein two amino substituents are selected from alkyl, aryl or aralkyl; alkanoylamino, aroylamino, aralkanoyl Amino, substituted alkanoylamino, substituted arylamino, substituted aralkanoylamino, thiol, alkylthio, arylthio, aralkylthio, alkylthiocarbonyl, aryl Alkylthiocarbonyl, aralkylthiocarbonyl, alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, sulfonamide, e.g. SO ₂ NH ₂ , substituted sulfonamide, nitro, cyano, carboxyl , carbamoyl such as CONH ₂ , substituted carbamoyl such as CONH alkyl, CONH aryl, CONH aralkyl or wherein on nitrogen there are two substituents selected from alkyl, aryl or aralkyl alkoxycarbonyl, aryl, substituted aryl, guanidino, heterocyclic groups such as indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidinyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, homopiperazinyl, etc., and substituted heterocyclic groups. Wherein the above substituents are further substituted by alkyl, alkoxy, aryl or aralkyl.

The term "halogen" or "halo" refers to fluorine, chlorine, bromine and iodine.

The term "aryl" means a monocyclic or bicyclic aromatic hydrocarbon group having 6 to 12 carbon atoms in the ring portion, such as phenyl, naphthyl, biphenyl and diphenyl, each of which may be substituted.

The term "aralkyl" refers to an aryl group bonded directly through an alkyl group or a substituted aryl group such as benzyl.

The term "aryloxy" refers to an aryl group or a substituted aryl group directly bonded through an alkoxy group such as methoxy or ethoxy.

The term "substituted aryl" refers to an aryl group substituted with, for example, one to four of the following substituents: such as alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted Alkynyl, aryl, substituted aryl, aralkyl, halo, trifluoromethoxy, trifluoromethyl, hydroxyl, alkoxy, alkanoyl, alkanoyloxy, aryloxy, aryl Alkoxy, amino, alkylamino, arylamino, aralkylamino, dialkylamino, alkanoylamino, thiol, alkylthio, ureido, nitro, cyano, carboxyl, carboxyalkyl, Carbamoyl, alkoxycarbonyl, alkylthiocarbonyl, arylthiocarbonyl, arylsulfonylamine, sulfonic acid, alkylsulfonyl, sulfonamido, aryloxy and the like. The substituent may be further substituted with hydroxy, halo, alkyl, alkoxy, alkenyl, alkynyl, aryl or aralkyl.

The term "heteroaryl" refers to an optionally substituted aryl, such as a 4 to 7 membered monocyclic, 7 to 11 membered bicyclic, or 10 to 15 membered tricyclic ring system, which has at least one heteroatom and A ring of at least one carbon atom, eg, pyridine, tetrazole, indazole.

The term "alkenyl" refers to a straight or branched chain hydrocarbon group of 2 to 20 carbon atoms, preferably 2 to 15 carbon atoms, and most preferably 2 to 8 carbon atoms, having one to four double bonds.

The term "substituted alkenyl" refers to alkenyl substituted by, for example, one to two of the following substituents: such as halo, hydroxy, alkoxy, alkanoyl, alkanoyloxy, amino, alkylamino, Dialkylamino, alkanoylamino, thiol, alkylthio, alkylthiocarbonyl, alkylsulfonyl, sulfonamide, nitro, cyano, carboxyl, carbamoyl, substituted carbamoyl, guanidine base, indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidinyl, pyridyl, pyrimidinyl, etc.

The term "alkynyl" refers to a straight or branched chain hydrocarbon group of 2 to 20 carbon atoms, preferably 2 to 15 carbon atoms, and most preferably 2 to 8 carbon atoms, having one to four triple bonds.

The term "substituted alkynyl" refers to an alkynyl group substituted by, for example, a substituent such as halo, hydroxy, alkoxy, alkanoyl, alkanoyloxy, amino, alkylamino, dialkylamino, Alkanoylamino, thiol, alkylthio, alkylthiocarbonyl, alkylsulfonyl, sulfonamide, nitro, cyano, carboxyl, carbamoyl, substituted carbamoyl, guanidino, and heterocyclyl , such as imidazolyl, furyl, thienyl, thiazolyl, pyrrolidinyl, pyridyl, pyrimidinyl and the like.

The term "cycloalkyl" refers to an optionally substituted saturated cyclic hydrocarbon ring system, preferably comprising 1 to 3 rings and 3 to 7 carbons per ring, which may be further combined with an unsaturated _C3 - _C7 carbocyclic ring fused. Exemplary groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, cyclododecyl, and adamantyl. Exemplary substituents include one or more of the alkyl groups described above, or one or more of the groups described above for the alkyl substituents.

The terms "heterocycle", "heterocyclic" and "heterocyclyl" refer to an optionally substituted fully saturated or unsaturated aromatic or non-aromatic ring group, for example, which is a 4 to 7 membered monocyclic ring, 7 to 11 membered bicyclic rings, or 10 to 15 membered tricyclic ring systems having at least one heteroatom in at least one ring containing carbon atoms. Each ring of the heterocyclic group containing heteroatoms can have 1, 2 or 3 heteroatoms selected from nitrogen atoms, oxygen atoms and sulfur atoms, wherein the nitrogen and sulfur heteroatoms can also be optionally oxidized, and the nitrogen heteroatoms can also be Can optionally be quaternized. A heterocyclyl group can be attached to any heteroatom or carbon atom.

唑烷基、异

唑啉基、异

唑基、噻唑基、噻二唑基、噻唑烷基、异噻唑基、异噻唑烷基、呋喃基、四氢呋喃基、噻吩基、

二唑基、哌啶基、哌嗪基、2-氧代哌嗪基、2-氧代哌啶基、高哌嗪基、2-氧代高哌嗪基、2-氧代吡咯烷基、2-氧杂氮杂基(oxazepinyl)、氮杂

基(azepinyl)、4-哌啶酮基、吡啶基、N-氧代-吡啶基、吡嗪基、嘧啶基、哒嗪基、四氢吡喃基、吗啉基、硫吗啉基、硫吗啉基亚砜(thiamorpholinyl sulfoxide)、硫吗啉基砜(thiamorpholinyl sulfone)、1，3-二氧戊环(dioxolane)和四氢-1，1-二氧代噻吩基、二

烷基、异噻唑烷基、硫杂环丁基(thietanyl)、硫杂环丙烷基(thiiranyl)、三嗪基和三唑基，等等。Exemplary monocyclic heterocyclyl groups include pyrrolidinyl, pyrrolyl, indolyl, pyrazolyl, oxetanyl, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, Azolyl,

Azolidinyl, iso

Azolinyl, iso

Azolyl, thiazolyl, thiadiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, furyl, tetrahydrofuryl, thienyl,

Diazolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, homopiperazinyl, 2-oxohomopiperazinyl, 2-oxopyrrolidinyl, 2-oxaazepine Base (oxazepinyl), aza

Base (azepinyl), 4-piperidinonyl, pyridyl, N-oxo-pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, sulfur Morpholinyl sulfoxide (thiamorpholinyl sulfoxide), thiamorpholinyl sulfone (thiamorpholinyl sulfone), 1,3-dioxolane (dioxolane) and tetrahydro-1,1-dioxothienyl, di

Alkyl, isothiazolidinyl, thietanyl, thiiranyl, triazinyl and triazolyl, and the like.

唑基、苯并噻吩基、奎宁环基、喹啉基、喹啉基-N-氧化物、四氢异喹啉基、异喹啉基、苯并咪唑基、苯并吡喃基、吲嗪基、苯并呋喃基、色酮基(chromonyl)、香豆素基(coumarinyl)、噌啉基、喹喔啉基、吲唑基、吡咯并吡啶基、呋喃并吡啶基(例如呋喃并[2，3-c]吡啶基、呋喃并[3，1-b]吡啶基]或呋喃并[2，3-b]吡啶基)、二氢异吲哚基、二氢喹唑啉基(例如3，4-二氢-4-氧代-喹唑啉基)、苯并异噻唑基、苯并异

唑基、苯并二嗪基(benzodiazinyl)、苯并呋咱基(benzofurazanyl)、苯并噻喃基、苯并三唑基、苯并吡唑基、二氢苯并呋喃基、二氢苯并噻吩基、二氢苯并噻喃基、二氢苯并噻喃基砜(dihydrobenzothiopyranyl sulfone)、二氢苯并吡喃基、二氢吲哚基、吲唑基、异色满基(isochromanyl)、异二氢氮杂茚基、1，5-二氮杂萘基(naphthyridinyl)、2，3-二氮杂萘基、胡椒基酰基、嘌呤基、吡啶并吡啶基、喹唑啉基、四氢喹啉基、噻吩并呋喃基、噻吩并吡啶基、噻吩并噻吩基等。Exemplary bicyclic heterocyclyl groups include 2,3-dihydro-2-oxo-1H-indolyl, benzothiazolyl, benzo

Azolyl, benzothienyl, quinuclidinyl, quinolinyl, quinolinyl-N-oxide, tetrahydroisoquinolyl, isoquinolyl, benzimidazolyl, benzopyranyl, indole Azinyl, benzofuryl, chromonyl (chromonyl), coumarinyl (coumarinyl), cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, furopyridyl (such as furo[ 2,3-c]pyridyl, furo[3,1-b]pyridyl] or furo[2,3-b]pyridyl), dihydroisoindolyl, dihydroquinazolinyl (e.g. 3,4-dihydro-4-oxo-quinazolinyl), benzisothiazolyl, benziso

Azolyl, benzodiazinyl, benzofurazanyl, benzothiopyranyl, benzotriazolyl, benzopyrazolyl, dihydrobenzofuranyl, dihydrobenzo Thienyl, dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone, dihydrobenzopyranyl, indolinyl, indazolyl, isochromanyl (isochromanyl), Isodihydroazindenyl, 1,5-naphthyridinyl, 2,3-naphthyridinyl, piperonyl acyl, purinyl, pyridopyridinyl, quinazolinyl, tetrahydro Quinolinyl, thienofuryl, thienopyridyl, thienothienyl and the like.

Exemplary substituents include one or more alkyl or aralkyl groups described above, or one or more groups described above as alkyl substituents. Also included are smaller heterocyclic groups such as epoxides and aziridines.

The term "carbocycle" refers to a stable saturated or partially unsaturated monocyclic hydrocarbon ring of 3 to 7 carbon atoms such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl. The term "optionally substituted" refers to "carbocycle" herein, which means that the carbocycle may be substituted at one or more substitutable ring positions by one or more groups independently selected from the group consisting of: alkane (preferably lower alkyl), alkoxy (preferably lower alkoxy), nitro, monoalkylamino (preferably lower alkylamino), dialkylamino (preferably di[lower] alkylamino), cyano radical, halo, haloalkyl (preferably trifluoromethyl), alkanoyl, aminocarbonyl, monoalkylaminocarbonyl, dialkylaminocarbonyl, alkylamido (preferably lower alkylamido), alkoxyalkane (preferably lower alkoxy[lower]alkyl), alkoxycarbonyl (preferably lower alkoxycarbonyl), alkylcarbonyloxy (preferably lower alkylcarbonyloxy) and aryl (preferably phenyl), The aryl is optionally substituted with halo, lower alkyl and lower alkoxy.

The term "heteroatom" shall include oxygen, sulfur and nitrogen.

The compounds of formula I may form salts which are also within the scope of this invention. Pharmaceutically acceptable (ie, non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful, for example, in isolating or purifying compounds of the invention.

The compounds of formula I can form salts with alkali metals such as sodium, potassium and lithium, with alkaline earth metals such as calcium and magnesium, with organic bases such as dicyclohexylamine, tributylamine, pyridine and amino acids such as arginine, lysine and the like. These salts can be formed according to methods known to those of ordinary skill in the art.

The compounds of formula I can form salts with various organic and inorganic acids. These salts include those formed with hydrogen chloride, hydrogen bromide, methanesulfonic acid, sulfuric acid, acetic acid, trifluoroacetic acid, oxalic acid, maleic acid, benzenesulfonic acid, toluenesulfonic acid and various other salts (e.g., Nitrates, Phosphates, Borates, Tartrates, Citrates, Succinates, Benzoates, Ascorbates, Salicylates, etc.). These salts can be formed according to methods known to those of ordinary skill in the art.

In addition, zwitterions ("inner salts") may be formed.

All stereoisomers of the compounds of the invention are contemplated in admixture or in pure or substantially pure form. The definition of compounds according to the invention includes all possible stereoisomers and mixtures thereof. It very particularly includes racemic forms and individual optical isomers which have specific activities. Racemic forms may be resolved by physical methods, for example by fractional crystallization, separation or crystallization of diastereoisomeric derivatives, or by separation by chiral column chromatography. These individual optical isomers can be obtained from racemates by conventional methods, for example, salt formation with an optically active acid, followed by crystallization.

Compounds of formula I may also be in the form of prodrugs. Any compound that will be converted in vivo to yield a biologically active agent (ie, a compound of formula I) is a prodrug within the scope and spirit of the invention.

Various forms of prodrugs are known in the art. For examples of these prodrug derivatives, see:

a) Design of Prodrugs , edited by H. Bundgaard, (Elsevier, 1985) and Methods in Enzymology , Vol. 42 , p.309-396, edited by K. Widder, et al. (Acamedic Press, 1985);

b) A Textbook of Drug Design and Development , edited by Krosgaard-Larsen and H. Bundgaard, Chapter 5, "Design and Application of Prodrugs," by H. Bundgaard, p.113-191 (1991); and

c) H. Bundgaard, Advanced Drug Delivery Reviews , 8 , 1-38 (1992).

It is further understood that solvates (eg hydrates) of compounds of formula I are also within the scope of the present invention. Methods of solvation are generally known in the art.

Practicality

The present invention is based on the discovery that certain pyrrolotriazines are inhibitors of protein kinases. In more detail, pyrrolotriazines such as those described in the present invention inhibit the protein tyrosine kinase activity of members of the HER family of receptors. These inhibitors would be useful in the treatment of proliferative diseases that depend on signaling from one or more of these receptors. These diseases include psoriasis, rheumatoid arthritis, and solid tumors of the lung, head and neck, breast, colon, ovary, and prostate. The present invention relates to a pharmaceutical composition of a compound of formula I, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier for treating hyperproliferative disorders in mammals. In particular, the pharmaceutical composition is expected to inhibit the growth of primary and recurrent solid tumors associated with HER1 (EGF receptor) and HER2, especially those tumors whose growth and spread are significantly dependent on HER1 or HER2, such as Includes bladder, squamous cell, head, colorectum, esophagus, gynecology (eg, ovary), pancreas, breast, prostate, vulva, skin, brain, genitourinary tract, lymphatic system (eg, thyroid), stomach, larynx, and lungs of cancer. In another specific embodiment, the compounds of the invention are also useful in the treatment of non-cancerous diseases such as psoriasis and rheumatoid arthritis.

Thus according to a further aspect of the present invention there is provided the use of a compound of formula I, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for producing an antiproliferative effect in a warm-blooded animal such as a human.

Thus according to a further aspect of the present invention there is provided a method of producing an antiproliferative effect in a warm-blooded animal (such as a human) in need of such treatment, said method comprising administering an effective amount of a compound of formula I as defined herein or its pharmaceutical The above acceptable salts are administered to the animals.

Due to their ability to inhibit HER1, HER2 and HER4 kinases, the compounds of the invention are useful in the treatment of proliferative diseases, including psoriasis and cancer. The HER1 receptor kinase has been shown to be expressed and activated in many solid tumors including head and neck, prostate, non-small cell lung, colorectal and breast cancers. Likewise, the HER2 receptor kinase has been shown to be overexpressed in breast, ovarian, lung and gastric cancers. Monoclonal antibodies that down-regulate the abundance of the HER2 receptor or inhibit signaling through the HER1 receptor have been shown to have anti-tumor efficacy in preclinical and clinical studies. It is therefore expected that inhibitors of HER1 and HER2 kinases will have efficacy in treating tumors that are dependent on signals from either receptor. In addition, these compounds would have efficacy in inhibiting tumors dependent on HER receptor heterodimer signaling. These compounds are contemplated as single agents or in combination with other chemotherapeutic agents such as Taxol , doxorubicin (adriamycin) and cisplatin combination (simultaneously or sequentially) has efficacy. Because HER1 and HER2 signaling have been shown to regulate the expression of angiogenic factors such as vascular endothelial growth factor (VEGF) and interleukin 8 (IL8), these compounds are expected to have inhibitory effects due to angiogenesis in addition to inhibition of tumor cell proliferation and survival. antitumor effect. The HER2 receptor has been shown to be involved in synoviocyte hyperproliferation in rheumatoid arthritis and may contribute to the angiogenic component of the inflammatory disease state. The inhibitors described in this invention are therefore expected to be effective in the treatment of rheumatoid arthritis. The ability of these compounds to inhibit HER1 further increases their utility as anti-angiogenic agents. See the following documents and references cited therein: Schlessinger J., "Cell signaling by receptortyrosine kinases", Cell 103(2), p.211-225 (2000); Cobleigh, MA, Vogel, CL, Tripathy, D ., Robert, NJ, Scholl, S., Fehrenbacher, L., Wolter, JM, Paton, V., Shak, S., Lieberman, G., and Slamon, DJ, "Multinational study of the efficacy and safety of humanized anti -HER2 monoclonal antibody in women who have HER2-overexpressing metastatic breast cancer that has progressed afterchemotherapy for metastatic disease", J.of Clin.Oncol.17(9), p.2639-2648(1999); Baselga, J., Pfister, D., Cooper, MR, Cohen, R., Burtness, B., Bos, M., D'Andrea, G., Seidman, A., Norton, L., Gunnett, K., Falcey, J., Anderson , V., Waksal, H., and Mendelsohn, J., "Phase I studies of anti-epidermal growth factor receptor chimeric antibody C225 alone and in combination with cisplatin", J. Clin. Oncol. 18(4), p.904 -914 (2000); Satoh, K., Kikuchi, S., Sekimata, M., Kabuyama, Y., Homma, MK, and Homma Y., "Involvement of ErbB-2 inrheumatoid synovial cell growth", Arthritis Rheum. 44(2), p.260-265(2001).

Anti-proliferative therapy as previously defined herein may be used as a sole therapy or may comprise, in addition to the compounds of the present invention, one or more other substances and/or treatments. Such conjoint treatment may be accomplished by simultaneous, sequential or separate administration of the individual components of the treatment. The compounds of the invention may also be used in combination with known anticancer and cytotoxic agents and treatments including radiation. If formulated as a fixed dose, these combination products employ the compounds of this invention within the dosage range described below and the other pharmaceutically active agent within its approved dosage range. Compounds of formula I may be used sequentially with known anticancer and cytotoxic agents and treatments including radiation when a combination formulation is inappropriate.

It is general practice in the field of medical oncology to use a combination of different forms of therapy to treat each cancer patient. In medical oncology, in addition to antiproliferative therapy as previously defined herein, the other component of the combination therapy may be: surgery, radiotherapy or chemotherapy. These chemotherapy can include three main classes of therapeutic agents:

(i) Anti-angiogenic agents acting by a mechanism different from the above definition (eg, linomide, inhibitors of integrin αvβ3 function, angiostatin, razoxane);

(贝伐单抗(bevacizurnab))和Erbitux(西妥昔单抗(cetuxirnab))；酪氨酸激酶抑制剂和丝氨酸/苏氨酸激酶抑制剂)；和(ii) Cytostatic agents such as antiestrogens (eg tamoxifen, toremifene, raloxifene, droloxifene ), indoxifene), progestogens (such as megestrol acetate), aromatase inhibitors (such as anastrozole, letrozole ), borazole, exemestane), antihormones, antiprogestogens, antiandrogens (such as flutamide, nilutamide (nilutamide, bicalutamide, cyproterone acetate), LHRH agonists and antagonists (eg, gosereline acetate, leuprolide), Inhibitors of testosterone 5α-dihydroreductase (such as finasteride (finasteride)), farnesyl transferase inhibitors, anti-invasion agents (such as metalloproteinase inhibitors such as marimastat ( marimastat) and inhibitors of urokinase plasminogen activator receptor function) and growth factor function, (these growth factors include, for example, EGF, FGF, platelet-derived growth factor and hepatocyte growth factor, these inhibitors Including growth factor antibodies, growth factor receptor antibodies such as Avastin

(bevacizumab) and Erbitux (cetuxirnab); tyrosine kinase inhibitors and serine/threonine kinase inhibitors); and

(紫杉醇(paclitaxel)、Taxotere(多西紫杉醇(docetaxel))和比较新的微管剂(microbtubule agents)例如epothilone类似物、discodermolide类似物和eleutherobin类似物)；拓扑异构酶(topoisomerase)抑制剂(例如表鬼臼毒素(epipodophyllotoxins)如依托泊苷(etoposide)和替尼泊苷(teniposide)、安吖啶(amsacrine)、托泊替康(topotecan)、伊立替康(irinotecan))；细胞周期抑制剂(例如flavopyridol)；生物反应调节物和蛋白酶体(proteasome)抑制剂例如Velcade

(bortezomib)。(iii) Antiproliferative/antineoplastic agents and combinations thereof, as used in medical oncology, such as antimetabolites (e.g. antifolates such as methotrexate, fluoropyrimidines such as 5- Fluorouracil, purine and adenosine analogues, cytosine arabinoside); intercalating antineoplastic antibiotics (e.g. anthracyclines such as doxorubicin, daunorubicin ), epirubicin (epirubicin) and idarubicin (idarubicin), mitomycin (mitomycin)-C, actinomycin D (dactinomycin), plicamycin (mithramycin)); platinum derivatives ( eg cisplatin, carboplatin); alkylating agents (eg nitrogen mustard, melphalan, chlorambucil, busulphan, cyclophosphamide, iso Cyclophosphamide, ifosfamide nitrosoureas, thiotepa; anti (mitotic) agents (eg vinca alkaloids like vincristine, vinorelbine (vinorelbine), vinblastine and vinflunine and taxoids such as Taxol

(paclitaxel, Taxotere (docetaxel) and newer microtubule agents such as epothilone analogs, discodermolide analogs and eleutherobin analogs); topoisomerase inhibitors (e.g. epipodophyllotoxins ) such as etoposide and teniposide, amsacrine, topotecan, irinotecan); cell cycle inhibitors (eg flavopyridol); biological Response modifiers and proteasome inhibitors such as Velcade

(bortezomib).

As mentioned above, the compounds of formula I according to the invention are of interest for their antiproliferative effect. These compounds of the invention are expected to be useful in a wide range of disease states including cancer, psoriasis and rheumatoid arthritis.

More specifically, compounds of Formula I are useful in the treatment of a variety of cancers, including (but not limited to) the following:

- Cancer, including bladder cancer, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer (including small cell lung cancer), esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, stomach cancer, cervical cancer, thyroid cancer, prostate cancer, and skin cancers including squamous cell carcinoma;

- Tumors of Mesenchymal origin, including fibrosarcoma and rhabdomyosarcoma;

- Tumors of the central and peripheral nervous systems, including astrocytomas, neuroblastomas, gliomas, and schwannomas; and

- Other tumors, including melanoma, seminoma, teratocarcinoma, and osteosarcoma.

In general due to the key role of kinases in the regulation of cell proliferation, inhibitors can act as reversible cytostatic agents useful in the treatment of any disease process characterized by abnormal cell proliferation, such as benign prostatic hyperplasia, familial adenomatosis Familial adenomatosis polyposis, neurofibromatosis, pulmonary fibrosis, arthritis, psoriasis, glomerulonephritis, restenosis after angioplasty or vascular surgery, hypertrophic scar formation, and inflammatory bowel disease sick.

Compounds of formula I are especially useful in the treatment of tumors in which tyrosine kinase activity is highly affected, such as colon, lung and pancreatic tumors. Tumor development in a mammalian host is reduced by administering a composition (or combination) of compounds of the invention.

The compounds of formula I are also useful in the treatment of diseases other than cancer which are associated with signal transduction pathways acting through growth factor receptors such as HER1 (EGF receptor), HER2 or HER4.

The pharmaceutical compositions of the invention comprising the active ingredient may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oleaginous suspensions, dispersible powders or granules, emulsions, hard or Soft capsule or syrup or elixir. Compositions intended for oral use may be prepared according to any methods known in the art for the manufacture of pharmaceutical compositions, and these compositions may contain one or more agents selected from sweetening agents, flavoring agents, coloring agents and preservatives so as to provide Pharmaceutically elegant and tasty preparation. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium or sodium phosphate; granulating and disintegrating agents such as microcrystalline cellulose, croscarmellose sodium, corn starch or alginic acid; binders such as starch, gelatin, polyvinylpyrrolidone or acacia, and lubricants such as magnesium stearate, stearic acid or talc. These tablets may be uncoated or they may be coated by known techniques to mask the unpleasant taste of the drug or to delay disintegration and absorption in the gastrointestinal tract, thereby providing sustained action over a longer period. For example, water soluble taste masking materials such as hydroxypropyl-methylcellulose or hydroxypropyl-cellulose, or time delay materials such as ethylcellulose, cellulose acetate buryrate may be employed.

Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is admixed with an inert solid diluent such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with a water-soluble carrier such as polyethylene glycol (polyethyleneglycol). ) or an oil medium such as peanut oil, liquid paraffin or olive oil.

Aqueous suspensions contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. These excipients are suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethyl-cellulose, sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and acacia Gum; dispersing or wetting agents may be naturally occurring phospholipids, such as lecithin, or condensation products of olefin oxides with fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide with long-chain aliphatic alcohols , such as heptadecaethylene-oxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitan monooleate , or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents and one or more sweetening agents agents such as sucrose, saccharin or aspartame.

Oleaginous suspensions may be formulated by suspending the active ingredient in a vegetable oil, such as arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. These oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those described above, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of antioxidants such as butylated hydroxyanisol or alpha-tocopherol.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Exemplary of suitable dispersing or wetting agents and suspending agents are those mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, may also be present. These compositions can be preserved by the addition of antioxidants such as ascorbic acid.

The pharmaceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, or a mineral oil, such as liquid paraffin, or mixtures of these. Suitable emulsifiers may be naturally occurring phospholipids such as soybean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides such as sorbitan monooleate, and the condensation of said partial esters with ethylene oxide Products such as polyoxyethylene sorbitan monooleate. These emulsions may also contain sweetening, flavoring, preservative and antioxidants.

Syrups and elixirs may be formulated with sweetening agents, such as glycerol, propylene glycol, sorbitol or sucrose. These formulations may also contain a demulcent, a preservative, flavoring and coloring agents and antioxidant.

Pharmaceutical compositions may be in the form of sterile injectable aqueous solutions. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution.

The sterile injectable preparation may also be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in the oily phase. For example, the active ingredient may first be dissolved in a mixture of soybean oil and lecithin. The oil solution is then introduced into a water and glycerin mixture and processed to form a microemulsion.

Injectable solutions or microemulsions can be introduced into the patient's bloodstream by local bolus injection. Alternatively, it may be advantageous to administer the solution or microemulsion in such a way as to maintain a constant circulating concentration of the compound of the invention. To maintain this constant concentration, a continuous intravenous delivery device can be utilized. An example of such a device is the Deltec CADD-PLUS.TM. model 5400 intravenous pump device.

These pharmaceutical compositions may be in the form of sterile injectable aqueous or oily suspensions for intramuscular and subcutaneous administration. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol. In addition, sterile fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

Compounds of formula I may also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights and fatty acid esters of polyethylene glycols.

For topical use, creams, ointments, gels, solutions or suspensions, etc., containing a compound of formula I are employed. (For purposes of this application, topical application will include mouthwashes and gargles).

The compounds of the present invention may be administered in intranasal form by topical use of appropriate intranasal vehicles and delivery devices, or by transdermal routes using transdermal skin patches well known to those of ordinary skill in the art. . To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen. The compounds of the invention may also be delivered in suppositories using bases such as cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights, and fatty acid esters of polyethylene glycols.

When the compounds according to the invention are administered to human patients, the daily dosage will normally be determined by the attending physician from the usual usual dosage which will vary according to the age, weight, sex and response of the individual patient, and the severity of the patient's symptoms.

If formulated as a fixed dose, these combination products employ the compounds of this invention within the dosage range described above and the other pharmaceutically active agent or treatment within its approved dosage range. Compounds of formula I may also be administered sequentially with known anticancer or cytotoxic agents when a combination formulation is inappropriate. The invention does not limit the order of administration; compounds of formula I may be administered before or after administration of known anticancer or cytotoxic agents.

The compounds may be administered in a single dose or in 2 to 4 divided doses in the dosage range of about 0.05 to 200 mg/kg/day, preferably less than 100 mg/kg/day.

biological test

HER1, HER2, or HER4 kinase assay

The compound of interest was mixed with 20mM Tris.HCl, pH7.5, 10mM MnCl ₂ , 0.5mM dithiothreitol, 0.1mg/ml bovine serum albumin, 0.1mg/ml poly(glu/tyr, 4:1), 1 μM ATP and 4 μCi/ml [γ- ³³ P]ATP in kinase buffer. Poly(glu/tyr, 4:1) is a synthetic polymer used as a phosphoryl acceptor and was purchased from Sigma Chemicals. The kinase reaction was initiated by adding enzyme and the reaction mixture was incubated at 26°C for 1 hour. The reaction was stopped by adding EDTA to 50 mM and the protein was precipitated by adding trichloroacetic acid to 5%. Precipitated protein was recovered by filtration on Packard Unifilter plates and the amount of bound radioactivity was measured in a Topcount scintillation counter.

For the production of recombinant HER1 and HER4, the cytoplasmic sequence of the receptor was expressed in insect cells as a GST fusion protein, which was purified by affinity chromatography. The cytoplasmic sequence of HER2 was subcloned into the baculovirus expression vector pBlueBac4 (Invitrogen) and expressed as an untagged protein in insect cells. The recombinant protein was partially purified by ion exchange chromatography.

Compounds of the invention inhibit HER1, HER2 and HER4 kinases with _IC50 values between 0.001 and 25 [mu]M. Preferred compounds have _IC50 values between 0.001-5.0 [mu]M. More preferred compounds have _IC50 values between 0.001-1.0 [mu]M. Most preferably compounds have _IC50 values between 0.001-0.1 [mu]M.

Preparation

In general, certain compounds of formula I can be prepared according to the following schemes and the knowledge of one of ordinary skill in the art. Additional preparative information can also be found in co-pending US Patent Application Serial No. 09/573,829 and International Publication No. WO 00/71129, filed May 18, 2000, both of which are incorporated herein by reference.

plan 1

where X=halogen, Y=N or O, alkyl= _CH3 or nBu

step 1

The first step in Scheme 1 is by treating compound i with a thiol such as methyl mercaptan or butanethiol or their sodium salts in an anhydrous solvent such as THF under an inert atmosphere such as _N2 (reference WO 03/042172A2) to This is accomplished by obtaining compound ii.

step 2

Halogenation of the 5-methyl group of compound ii is achieved by treatment with a halogenating reagent such as N-bromosuccinimide. The reaction is carried out under an inert atmosphere such as Ar gas in the presence of a catalyst such as dibenzoyl peroxide or 2,2'-azobisisobutyronitrile and yields 5-halomethyl-pyrrolotriazine compound iii .

step 3

Treatment of compound iii with a primary or secondary amine or alcohol in the presence of a base such as NaHCO or _{triethylamine} or diisopropylethylamine in a solvent such as acetonitrile or N,N-dimethylformamide affords the intermediate Compound iv.

step 4

Treatment of intermediate compound iv with aniline in the presence of _HgCl2 in a solvent such as toluene affords 4-substituted pyrrolotriazine compound v.

step 5

Alternatively compounds of formula iv can be treated with a suitable oxidizing agent such as m-chloroperbenzoic acid in _a solvent such as _CH2Cl2 to afford sulfones vi.

step 6

Sulfones vi can be converted to compounds v by treatment with primary or secondary amines or alcohols in _an inert solvent such as _CH2Cl2 .

Alternatively, compounds of general formula I can be prepared as shown in Scheme 2.

Scenario 2

where X=halogen, Y=N or O

step 1

The first step of Scheme 2 is accomplished by treating compound i with a halogenating reagent such as N-bromosuccinimide under an inert atmosphere such as Ar (cf. WO 03/042172 A2). The reaction is carried out in a suitable solvent such as _CCl4 in the presence of a catalyst such as dibenzoyl peroxide or 2,2'-azobisisobutyronitrile to give dihalopyrrolotriazine compounds vii.

step 2

Compound vii can be converted to the ammonium salt compound viii by treatment with a tertiary base such as triethylamine in anhydrous solvent such as THF.

step 3

Treatment of compound viii with an amine or its anion in an anhydrous solvent such as acetonitrile, chloroform or THF affords ammonium salt compound ix.

step 4

Conversion of compound ix to pyrrolotriazine compound v can be accomplished by treating compound ix with a primary or secondary amine or alcohol in a solvent such as acetonitrile in the presence of a base such as diisopropylethylamine.

烷溶液处理或通过用三氟乙酸处理化合物的CH₂Cl₂溶液除去保护基而进一步改性制备得到游离胺xi。进一步的改性可在步骤2中通过在还原剂例如三乙酰氧基硼氢化钠存在下，在溶剂例如CH₂Cl₂中用羰基化合物例如丙醛处理化合物xi以得到被取代的胺xii而完成。Compounds of general formula x in Scheme 3 prepared by the above method, wherein the 5-methyl substituent contains a protecting group such as tert-butoxycarbonyl, can be obtained in step 1 by diethyl ether or 1,4 with anhydrous HCl -two

The free amines xi were prepared by treatment with trifluoroacetic acid in CH2Cl2 or by treatment of the compound _{in CH2Cl2} to remove the protecting group. Further modification can be accomplished in step 2 by treating compound xi with a carbonyl compound such as _{propionaldehyde} in the presence of a reducing agent such as sodium triacetoxyborohydride in a solvent such as _CH2Cl2 to give the substituted amine xii .

Option 3

Further compounds can be prepared as shown in Scheme 4. In step 1, compound xii can be obtained by treating compound ix with a secondary amine such as bis-(2-chloroethyl)amine in the presence of a base such as diisopropylethylamine in a suitable solvent such as acetonitrile. Compound xii can be further treated with a nucleophile such as hydrazine in step 2 to give compound xiii.

Option 4

Additional 5-substituted pyrrolotriazines can be prepared according to Scheme 5.

Option 5

step 1

Compound i can be treated with two equivalents of a brominating reagent such as N-bromosuccinimide in a solvent such as _CCl4 at elevated temperature. The resulting 5-dibromopyrrolotriazine can be converted to the corresponding dimethylacetal using methanol in the presence of a base such as NaHCO ₃ and then treated intermediately with an acid such as trifluoroacetic acid in the presence of water. Convert acetaldehyde to aldehyde compound xiv.

step 2

Treatment of aldehyde compound xiv with an organometallic reagent such as Grignard's reagent in anhydrous solvent such as THF affords alcohol xv.

step 3

Alcohols xv can be treated with primary or secondary amines or alcohols in the presence of a base such as _NaHCO3 in a suitable solvent such as acetonitrile to give compounds of formula xvi.

In addition, other compounds of formula I can be prepared using procedures generally known to those of ordinary skill in the art. In particular, the following examples provide additional methods for preparing compounds of the invention.

The invention will now be further described by the following working examples, which are preferred embodiments of the invention. All temperatures are in degrees Celsius (°C) unless otherwise indicated. "HPLC retention time" is the HPLC retention time obtained under the following conditions: column type and length, gradient time [unless otherwise indicated, all gradients are in 100% solvent A (10% MeOH, 90% _H2O , 0.1 %TFA) and end with 100% solvent B (90% MeOH, 10% _H2O , 0.1% TFA)], flow rate (mL/min). UV detection was always performed at 22OnM. These examples are illustrative rather than limiting, and it is understood that there may be other specific embodiments that fall within the spirit and scope of the invention as defined by the appended claims.

Example 1

5-[(4-amino-1-piperidinyl)methyl]-N-(3-chloro-4-fluorophenyl)pyrrolo[2,1-f][1,2,4]triazine- 4-amine

1A. Preparation of 5-methyl-4-methylthio-pyrrolo[2,1-f][1,2,4]triazine

NaSMe (1.85 g, 26.3 mmol) was added to 4-chloro-5-methyl-pyrrolo[2,1-f][1,2,4] sparged with _N2 at 0 °C Triazine (4.02 g, 24.0 mmol) (reference WO 03/042172A2) in solution in anhydrous THF (200 mL). Continue bubbling for 5 minutes. The reaction mixture was then stirred overnight at room temperature and concentrated in vacuo to leave a volume of about 50 mL. Dilute with _H2O (280 mL) and stir at 0 °C. The solid was filtered, washed with cold water and dried to give 1A (3.91 g, 91%). It had an analytical HPLC retention time = 3.38 min. (YMC S5 ODS column 4.6 x 50 mm, 10-90% aqueous methanol containing 0.2% phosphoric acid over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +1 = 180.

1B.[1-(4-Methylthio-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethyl)-piperidin-4-yl]-carbamic acid tert- Preparation of butyl ester

A mixture of 1A (1.94 g, 10.8 mmol), dibenzoyl peroxide (0.262 g, 1.08 mmol), NBS (2.12 g, 11.90 mmol) in _CCl (100 mL) was bubbled with N and then _immediately Heat at 85°C for 1.5 hours. The mixture was cooled to room temperature, and the precipitate was filtered off. The filtrate was concentrated in vacuo, diluted with dichloroethane (35 mL), and DIEA (2.24 mL, 12.96 mmol) and piperidin-4-yl-carbamate tert-butyl ester (2.38 g, 11.90 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. The mixture was diluted with saturated NaHCO ₃ (70 mL), and extracted with EtOAc (3×100 mL). The combined EtOAc extracts were washed with brine (1 x 100 mL), dried ( _MgSO4 ), filtered and concentrated in vacuo. The residue was purified by silica gel flash column to give 1B ₍ 2.87 g, 70%) (0.1%-2% MeOH- _CH2Cl2 ). It had an analytical HPLC retention time = 2.12 min. (YMC S5 ODS column 4.6 x 50 mm, 10-90% aqueous methanol containing 0.2% phosphoric acid over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +1 = 378.

1C. Preparation of 5-bromomethyl-4-chloro-pyrrolo[2,1-f][1,2,4]triazine

4-Chloro-5-methyl-pyrrolo[2,1-f][1,2,4]triazine (2.0g, 11.93mmol) (refer to WO03/042172 A2) and AIBN (195mg, 1.19mmol) The mixture in _CCl4 (80 mL) was heated at 100 °C under _N2 for 5 min and NBS (2.55 g, 14.3 mmol) was added. The reaction mixture was stirred for 10 minutes, then cooled to room temperature and filtered. The CCl ₄ layer was washed with dilute aqueous NaHCO ₃ , dried (MgSO ₄ ), filtered and concentrated to give 1C (2.70 g, 92%).

1D. Preparation of (4-chloro-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethyl)-triethyl-ammonium bromide

A mixture of 1C (2.7 g, 11 mmol), _Et3N (5 mL, 36 mmol) in THF (20 mL) was stirred at room temperature for 12 hours. The solid was filtered and rinsed with THF and _Et2O , dried to give 1D (3.38g, 89%). It had an analytical HPLC retention time = 0.776 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ = 267 .

1E. [4-(3-Chloro-4-fluoro-phenylamino)-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethyl]-triethyl-ammonium Preparation of bromide hydrochloride

A mixture of 1C (1.0 g, 2.2 mmol) and 3-chloro4-fluoro-aniline (418 mg, 2.87 mmol) in _CHCl3 (10 mL) was heated at 50 °C for 2 hours. The solid was filtered and rinsed with CHCl ₃ and dried to give 1E (1.24 g, 87.4%). It had analytical HPLC retention time = 2.19 min (ChromolithSpeedROD 4.6 x 50 mm, 10-90% methanol in water with 0.1% TFA over 4 min, 4 mL/min, monitored at 220 nm) and LC/MS M ⁺ = 376.

1 F.{1-[4-(3-Chloro-4-fluoro-phenylamino)-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethyl]-piper Preparation of pyridin-4-yl}-carbamic acid tert-T ester

method one:

A mixture of 1B (30 mg, 0.08 mmol), 3-chloro-4-fluoro-aniline (11 mg, 0.08 mmol) and _HgCl2 (24 mg, 0.088 mmol) in toluene (2 mL) was heated at reflux for 8 hours. Cool to room temperature, dilute with EtOAc (5 mL) and filter. The filtrate was concentrated, and the residue was purified by preparative HPLC to give oil IF.

Method Two:

A mixture of 1E (9.1 g, 18.4 mmol) and DIPEA (3.2 mL, 18.4 mmol) in _CH3CN (40 mL) was added dropwise to piperidin-4-yl- A suspension of tert-butyl carbamate (4.1 g, 20.3 mmol) in _CH3CN (55 mL). The reaction mixture was stirred at 70 °C for 1 h, then cooled to room temperature, after which _H2O (155 mL) was added slowly. The solid was filtered and rinsed with 15% _CH3CN / _H2O , then _H2O , and dried under vacuum to give 1F (7.84 g, 90%). It has analytical HPLC retention time = 2.73 min (ChromolithSpeedROD 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 mL/min, monitored at 220 nm) and LC/MS M ⁺ +1 = 475.

1G.5-[(4-amino-1-piperidinyl)methyl]-N-(3-chloro-4-fluorophenyl)pyrrolo[2,1-f][1,2,4]tri Preparation of oxazin-4-amine

Compound IF (from method one) was treated with 20% TFA/ _CH2Cl2 (3 mL) at ₀ °C, then stirred at room temperature for 2 hours. The reaction mixture was concentrated and purified by preparative HPLC to give the product as a TFA salt, which was treated with saturated NaHCO ₃ to give the free base 1G (4 mg, 13% for two steps). It has analytical HPLC retention time = 1.49 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 mL/min, monitored at 220 nm) and LC/MS M ⁺ +1 =375.

Example 2

5-[(4-Amino-1-piperidinyl)methyl]-N-4-pyridylpyrrolo[2,1-f][1,2,4]triazin-4-amine

NaHMDS in THF (722 μL, 0.722 mmol) was added to a mixture of pyridin-4-ylamine (34 mg, 0.361 mmol) in THF (500 μL). The mixture was cooled to 0° C. and a suspension of 1D (125 mg, 0.27 mmol) in DMF (800 μl) was added. The mixture was stirred at this temperature for 0.5 hours, and piperidin-4-yl-carbamate tert-butyl ester (144 mg, 0.72 mmol) was added to the cold mixture. The reaction mixture was heated at 50°C for 10 minutes and concentrated to remove THF. TFA (1 mL) was added and the mixture was stirred until the protecting group was removed (2 hours) (progress monitored by HPLC). TFA was removed in vacuo and saturated _NaHCO3 was added. The mixture was extracted with EtOAc, and the combined extracts were dried, concentrated, and triturated first with _Et2O to give the title compound (46 mg, 53%). Analytical HPLC retention time = 0.51 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +1 = 324 .

Example 3-37

Compounds 3-37 were prepared using methods similar to compounds in Example 2 using the corresponding amines.

Examples 38-121

method one:

Compounds (with HPLC annotation (a)) were prepared by the following standard methods.

In a dram vial was added 1D (55.0 mg, 0.16 mmol), aniline (0.16 mmol, 1.0 eq) and _CH3CN (1 mL). The mixture was shaken overnight at 65°C. To this mixture was added piperidin-4-yl-carbamate tert-butyl ester (34.9 mg, 0.17 mmol) followed by DIEA (28 μL, 16 mmol). The reaction was continued at 65°C for 3 hours. The mixture was concentrated; the residue was purified by preparative HPLC and the desired fractions were collected and concentrated. The resulting residue was dried overnight under high vacuum.

_CH2Cl2 (1.5 mL) and TFA ₍ 0.2 mL) were added to the above residue, and the reaction mixture was shaken at room temperature for 2 hours. The mixture was concentrated and dried overnight in speed vacuum to give a solid product. Preparative HPLC was used further only when the solid was impure.

Method Two:

Compounds (with HPLC symbol (b)) were prepared by the following standard methods.

A mixture of 1D (75 mg, 0.216 mmol) and aniline (1.0 equiv, 0.216 mmol) in N,N-dimethylacetamide (0.5 mL) was heated in a vial at 70 °C for 3-5 hours until obtaining Clear solution. The progress of the reaction was followed using HPLC. The reaction mixture was cooled to room temperature and piperidin-4-yl-carbamate tert-butyl ester (43 mg, 0.216 mmol) was added followed by N,N-diisopropylethylamine (75 μl). The reaction mixture was again heated at 70 °C overnight. _Once cooled, the reaction mixture was diluted with _CH2Cl2 (0.5 mL) and cooled to 0 °C. TFA (1.0 mL) was added and the mixture was stirred at ambient temperature overnight. The solvent was removed under reduced pressure (speedVac) and the residue was dissolved in methanol and purified by prep HPLC to give the desired product.

HPLC conditions:

(a): (YMC S5 ODS column 4.6×50 mm, 1 0-90% methanol aqueous solution, containing 0.2% H ₃ PO ₄ , lasted 4 minutes, 3 ml/min, monitored at 220nm)

(b): (Chromolith SpeedROD 4.6×50 mm, 1 0-90% methanol in water, containing 0.1% TFA, over 4 minutes, 4 ml/min, monitored at 220nm)

Note: Examples 97-99, 101 and 104 have been removed from the table due to duplication.

Examples 122-132

Compounds 122-132 were prepared from Compound 1D, 3-chloro-4-fluoroaniline and the corresponding amine or Boc protected amine by a route similar to that used for the preparation of Compound 38-121.

Example 133

5-[(4-amino-1-piperazinyl)methyl]-N-(3-chloro-4-fluorophenyl)pyrrolo[2,1-f][1,2,4]triazine- 4-amine

133A(5-{[bis-(2-chloro-ethyl)-amino]-methyl}-pyrrolo[2,1-f][1,2,4]triazin-4-yl)-(3 Preparation of -chloro-4-fluoro-phenyl)-amine

A mixture of compound 1E (50 mg, 0.1 mmol), bis-(2-chloroethyl)amine hydrochloride (18 mg, 0.1 mmol), DIEA (36 μl, 0.2 mmol) in CH ₃ CN (0.5 mL) Heat at 60°C for 3 hours. The mixture was cooled to room temperature and concentrated to give compound 133A, which was used directly in the next step. 133A has analytical HPLC retention time = 2.986 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +1 =416.

The crude 133A from _the last step was dissolved in pure anhydrous _N2H4 (0.5 mL) and heated at 100 °C for several hours. The mixture was cooled _to room temperature, diluted with _H2O and extracted with _CH2Cl2 . The combined extracts were washed with _brine , dried over _Na2SO4 , and concentrated in vacuo. The residue was purified by preparative HPLC to afford compound 133 (38.8 mg, ₁₀₀ % for two steps) after neutralization and extraction (with _CH2Cl2 ). Analytical HPLC retention time = 1.709 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 mL/min, monitored at 220 nm) and LC/MS M ⁺ +1 = 376.

Example 134

(3-Chloro-4-fluoro-phenyl)-[5-(morpholin-2-ylmethoxymethyl)-pyrrolo[2,1-f][1,2,4]triazine-4 -yl]-amine

134A 2-(4-methylthio-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethoxymethyl)-morpholine-4-carboxylic acid tert-butyl ester preparation of

A solution of compound 1A (1.0 g, 5.6 mmol) in _CCl4 (50 mL) was purged with nitrogen for one hour. Dibenzoyl peroxide (270 mg, 1.12 mmol) was added and the reaction mixture was heated to 86°C. N-Bromosuccinimide (1.04 g, 5.88 mmol) was added in one portion. After 30 minutes, the reaction was cooled to room temperature and filtered. The filtrate was concentrated, redissolved in toluene (10 mL) and treated with tert-butyl 2-hydroxymethyl-morpholine-4-carboxylate (1.5 g, 6.9 mmol). The solution was heated at 110°C for 8 hours, cooled to room temperature and concentrated. Flash chromatography on silica (20% EtOAc/hexanes) gave the product as a pale yellow oil which crystallized on standing (770 mg, 32%). HPLC _tR = 3.783 min (YMC S5 ODS 4.6 x 50 mm, 10-90% methanol in water, 4 min gradient, monitored at 220 nm). LC/MS (M+H)=178.

134B 2-[4-(3-Chloro-4-fluoro-phenylamino)-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethoxymethyl]-mol Preparation of tert-butyl morphine-4-carboxylate

2-(4-Methylthio-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethoxymethyl)-morpholine-4-carboxylic acid tert-butyl ester (60 mg, 0.15 mmol) in _CH2Cl2 (3 mL) was cooled _to 0 °C and treated with mCPBA (56 mg, 0.32 mmol ₎ in _CH2Cl2 (2 mL). The reaction was stirred at 0 °C for 15 minutes and then allowed to warm to room temperature. 3-Chloro-4-fluoroaniline was added to this solution and stirred at room temperature for one hour. The resulting orange solution was diluted with _CH2Cl2 and washed with saturated aqueous _NaHCO3 , then _saturated aqueous NaCl. _The organic layer was dried ( _Na2SO4 ), filtered and concentrated. Preparative reverse phase HPLC gave the desired compound (30 mg, 41%). HPLC _tR = 4.383 min (YMC S5ODS 4.6 x 50 mm, 10-90% methanol in water, 4 min gradient, monitored at 220 nm). LC/MS (M+H)=492.

A solution of 134B (30 mg, 0.06 mmol) in _CH2Cl2 (3 mL) was treated with trifluoroacetic acid (0.3 mL) at 0 ° _C by dropwise addition. The reaction _was stirred for two hours, then diluted with _CH2Cl2 and washed with saturated aqueous _NaHCO3 . _The organic layer was separated, dried ( _Na2SO4 ), filtered and concentrated. The crude compound was purified by _radial chromatography (1 mm plate, ₁₅ % MeOH/ _CH2Cl2 to 30% MeOH/ _CH2Cl2 ) to afford compound 134 (17 mg, 67%). HPLC _tR = 2.83 min (YMC S5 ODS 4.6 x 50 mm, 10-90% methanol in water, 4 min gradient, monitored at 220 nm). LC/MS (M+H)=392.

Example 135

4-amino-1-[[4-[(3-chloro-4-fluorophenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl]methyl] -(3R,4R)-rel-3-piperidinol

Compound 135A:

To a solution _of 1,2,3,6-tetrahydropyridine (1.66 g, 20.0 mmol) in anhydrous _CH2Cl2 (10 mL) was added triethylamine (3.35 mL, 24.0 mmol) followed by N- (Benzyloxycarbonyloxy)succinimide (5.23 g, 21.0 mmol) in anhydrous _CH2Cl2 (10 mL) _. The reaction mixture was stirred overnight at room temperature. The reaction mixture was diluted with _CH2Cl2 (50 mL), washed with 10% citric acid, saturated _{NaHCO3 ,} brine, and dried over _{anhydrous Na2SO4} . Concentration under reduced pressure gave 4.34 g of Compound 135A as an oil: (100%). Analytical HPLC retention time = 2.996 min (ChromolithSpeedROD column 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 mL/min, monitored at 254 nm).

¹ H-NMR (CDCl3): 7.20-7.35 (m, 5H), 5.88 (bs, 1H), 5.60-5.78 (m, 1H), 5.18 (s, 2H), 3.99 (t, J=2.64, 2H) , 3.59(t, J=5.69, 2H), 2.18(m, 2H).

Compound 135B

To a solution of compound 135A (1.1 g, 5.0 mmol) in anhydrous CH ₂ Cl ₂ (10 mL) cooled to 0° C. was added 75% m-CPBA (1.38 g, 6.0 mmol) in anhydrous CH ₂ Cl ₂ (5 ml) solution. The reaction mixture was stirred at 0°C for 15 minutes, then at room temperature for 3 hours. The reaction mixture was diluted with CH ₂ Cl ₂ (20 mL), and washed with sat. Na ₂ S ₂ O ₃ , sat. NaHCO ₃ , brine, and dried over anhydrous Na ₂ SO ₄ . Concentration under reduced pressure afforded 1.14 g (98%) of compound 135B as an oil. Analytical HPLC retention time = 2.279 min (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 mL/min, monitored at 254 nm).

¹ H-NMR (CDCl ₃ ): 7.20-7.36 (m, 5H), 5.05 (s, 2H), 3.80-3.96 (m, 1H), 3.70 (m, 1H), 3.47 (m, 1H), 3.22 ( bs, 1H), 3.07-3.20(m, 2H), 2.00(m, 1H), 1.87(m, 1H).

Compounds 135C and 135D:

A solution of sodium azide (100 mg, 1.5 mmol) in a 2:1 mixture of acetone-water (2 mL) was added to a solution of compound 135B (233 mg, 1.0 mmol) in anhydrous DMF (2 mL). The reaction mixture was heated at 80 °C overnight. The solvent was removed under reduced pressure, and the residue was dissolved in EtOAc (20 mL), washed with water, 10% LiCl and brine, and dried over _{anhydrous Na2SO4} . Concentration under reduced pressure gave an oil. Flash chromatography on silica gel (hexane-ethyl acetate: 8:2 to 7:3) gave 180 mg of compound 135C (early eluent, major isomer) as an oil and 98 mg as an oil Compound 135D (late eluent, minor isomer).

Compound 135C: ¹ H-NMR (CDCl ₃ ): 7.28-7.40 (m, 5H), 5.10 (s, 2H), 4.14 (dd, J1=4.03, J2=13.44, 1H), 4.02 (m, 1H), 3.50(m, 1H), 3.38(m, 1H), 3.00(m, 1H), 2.88(m, 1H), 2.70 and 2.40(part of m, 1H), 2.00(m, 1H), 1.50(m, 1H).

Compound 135D: ¹ H-NMR (CDCl ₃ ): 7.20-7.35 (m, 5H), 5.06 (s, 2H), 4.25 and 4.10 (part m, 1H), 3.99 (d, J=13.44, 1H), 3.50(m, 1H), 3.22(m, 1H), 2.85(t, J)=2.69, 1H), 2.73(m, 1H), 2.40(m, 1H), 1.90(m, 1H), 1.45(m , 1H).

Compound 135E

Water (0.05 mL) and triphenylphosphine (340 mg, 1.3 mmol) were added to a solution of compound 135C (180 mg, 0.65 mmol) in THF (5 mL), and the reaction mixture was heated to reflux for 6 hours. After cooling to room temperature, EtOAc (20 mL) was added to the reaction mixture. The organic layer was extracted with 1.0N HCl (10 mL x 2), and the combined aqueous layers were backwashed once with EtOAc (5 mL). 1.0 N NaOH was added to the aqueous layer to make it pH 10.0 and the mixture was extracted with EtOAc (20 mL x 2). The combined organic layers were dried over _{anhydrous Na2SO4} . Concentration under reduced pressure gave 165 mg of the amine intermediate as a colorless oil.

Triethylamine (0.11 mL, 0.78 mmol) followed by _Boc2O (156 mg, 0.72 mmol) was added to _a solution of 165 mg of the amine intermediate in anhydrous _CH2Cl2 (4 mL). The mixture was stirred overnight at room temperature. The reaction mixture was diluted with _CH2Cl2 , washed _with saturated _NaHCO3 , and dried over _{anhydrous Na2SO4} . Purification by silica gel flash chromatography (Hexane-EtOAc: 9:1 to 8:2) afforded 170 mg of compound 135E as a white solid. Analytical HPLC retention time = 2.859 minutes (ChromolithSpeedROD column 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 minutes, 4 ml/min, monitored at 254 nm) and

a LC/MS M ⁺ +1 = 351 ^{+ .1} H-NMR (CDCl ₃ ): 7.29-7.40 (m, 5H), 5.10 (s, 2H), 4.61 (bs, 1H), 4.32 (bs, 1H) , 3.90-4.30(m, 1H), 3.30-3.60(m, 2H), 2.80(m, 1H), 2.66(m, 1H), 1.90(m, 1H), 1.45(s, 9H), 1.40(m , 1H).

Compound 135F

A solution of compound 135E (170 mg) in 5 mL MeOH containing 10 mg Pd(OH) ₂ was stirred under hydrogen atmosphere (balloon) overnight. The catalyst was removed by filtration and rinsed with MeOH. The combined filtrates were concentrated under reduced pressure to afford 138 mg of compound 135F as an oil. Analytical HPLC retention time = 1.270 min (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% aqueous methanol containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +1 = ²¹⁷⁺ .

Compound 135G

Compound 135G was prepared from Compound 135D by a procedure similar to Compound 135E. Analytical HPLC retention time = 2.849 min (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 mL/min, monitored at 254 nm).

¹ H-NMR (CDCl ₃ ): 7.4O-7.52(m, 5H), 5.20(s, 2H), 4.30(m, 2H), 3.40(m, 1H), 2.95(m, 1H), 2.67(m , 2H), 2.08(m, 1H), 1.45-1.96(m, 3H), 1.45(s, 9H).

Compound 135H

Compound 135H was prepared from Compound 135G by a procedure similar to Compound 135F. Analytical HPLC retention time = 1.380 min (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 mL/min, monitored at 220 nm).

Compound 135

Compound 135 was prepared by a method similar to Example 1 using Compounds 135F and 1E. Compound 135 is a compound with analytical HPLC retention time = 1.666 minutes (ChromolithSpeedROD column 4.6 × 50 mm, 10-90% aqueous methanol, containing 0.1% TFA, experienced 4 minutes, 4 ml/min, monitored at 254nm) and LC/MS M ⁺ +1 = 391 ⁺ solid.

¹ H-NMR (CDCl ₃ ): 11.62 (s, 1H), 7.94 (s, 1H), 7.89 dd, J ₁ =2.60, J ₂ =6.61, 1H), 7.48 (d, J = 2.60, 1H), 7.45(m, 1H), 7.15(t, J=8.72, 1H), 6.51(d, J=2.60, 1H), 3.82(AB, J=13.60, Δv=26.94, 2H), 3.33(m, 1H) , 3.25(m, 1H), 3.08(d, J=12.09, 1H), 2.57(m, 1H), 2.22(t, J=12.03, 1H), 2.05(m, 1H), 1.97(m, 1H) , 1.43(m, 1H).

Alternatively, compound 135 can be prepared as shown below.

Preparation of Compound 135J

Compound 135J was prepared according to the published literature procedure: Jacob Szmuszkovicz et al., Heterocycles, 1994, 39(1), 163-170.

Preparation of Compound 135K:

Compound 135K was prepared according to the published literature procedure: Jacob Szmuszkovicz et al., Heterocycles, 1994, 39(1), 163-170.

Preparation of compound 135L:

Compound 135L was prepared from Compound 135K in a similar manner to Compound 1C. Analytical HPL retention time = 2.323 min (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 mL/min, monitored at 220 nm).

¹ H-NMR (CDCl3): 4.11 (dd, J1 = 3.09, J2 = 13.29, 1H), 3.95 (m, 1H), 3.50 (m, 1H), 3.38 (m, 1H), 2.90 (m, 1H) , 2.79(dd, J1=9.27, J2=13.29, 1H), 2.45(m, 1H), 2.00(m, 1H), 1.55(m, 1H), 1.46(s, 9H).

Preparation of Compound 135M:

Trifluoroacetic acid (5 mL) was added to a solution of compound 135 _L (0.6 g, 2.48 mmol) in anhydrous _CH2Cl2 cooled at 0 °C. The reaction mixture was stirred at 0 °C for 15 minutes, then allowed to warm to room temperature and stirred for 3 hours. _The solvent and TFA were removed under reduced pressure, and the residue was dissolved in _CH2Cl2 (20 mL). The organic layer was washed with saturated NaHCO ₃ , and the aqueous layer was supersaturated with solid NaCl and back extracted with EtOAc (15 mL×10). The combined organic extracts were dried over _{anhydrous Na2SO4} . Concentration in vacuo gave 350 mg of compound 135M as an oil. ¹ H-NMR (CDCl ₃ +CD ₃ OD): 3.55 (m, 1H), 3.43 (m, 1H), 3.18 (dd, J1=3.95, J2=12.63, 1H), 3.07 (d oft, J1=12.90 , J2=4.78, 1H), 2.74(m, 1H), 2.63(dd, J1=8.28, J2=12.58, 1H), 2.10(m, 1H), 1.57(m, 1H).

Preparation of compound 135N:

Compound 135N was prepared starting from compound 135M and 1E of Example 1 in a similar manner to compound 1F in Example 1 (using Method 2). Compound 135N was a solid with an analytical HPLC retention time = 2.099 minutes (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 minutes, 4 mL/min, monitored at 254 nm) and LC/ MS M ⁺⁺ 1=417 ⁺ .

Triphenylphosphine (262 mg, 1.0 mmol) was added to a solution of compound 135N (0.5 mmol) prepared above in a mixture of THF (5 mL) and water (0.05 mL). The reaction mixture was heated to reflux for 8 hours. After cooling to room temperature, the solvent was evaporated under reduced pressure and the residue was directly purified by flash chromatography on silica gel ( _{CH2Cl2 -} MeOH- _NH4OH : 95:5:0.5) to give 166 mg of the compound as a solid 135.

Example 136

3-amino-1-[[4-[(3-chloro-4-fluorophenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl]methyl] -(3R,4R)-rel-4-piperidinol

Compound 136 was prepared in a similar manner to Example 1 using Compounds 135H and 1E. Compound 136 was a solid with analytical HPLC retention time = 1.953 min (ChromolithSpeedROD column 4.6 x 50 mm, 10-90% aqueous methanol containing 0.1% TFA over 4 min, 4 mL/min, monitored at 254 nm) and LC/MS M ⁺ +1 = 391 ⁺ .

Example 137

4-amino-1-[[4-[(3-chloro-4-fluorophenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl]methyl] -(3R,4R)-(+)-rel-3-piperidinol

and

Example 138

4-amino-1-[[4-[(3-chloro-4-fluorophenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl]methyl] -(3R,4R)-(-)-rel-3-piperidinol

Racemic compound 135 was resolved using normal-phase chiral preparative HPLC (Chiralpak AD) using hexane-isopropanol-diethylamine (80:20:0.05) as mobile phase. Compound 137 (enantiomer A) and compound 138 (enantiomer B) were obtained as single enantiomers with >99% ee.

Example 139

N-(3-chloro-4-fluorophenyl)-5-[[4-[(2,2-dimethylpropyl)amino]-1-piperidinyl]methyl]pyrrolo[2,1 -f][1,2,4]triazin-4-amine

Glacial acetic acid (0.05 ml), followed by 3,3-dimethylbutyraldehyde (0.008 ml, 0.073 mmol) and sodium triacetoxyborohydride (25 mg, 0.12 mol) were added to the compound of Example 1 (19 mg, 0.05 mmol) in _{CH2Cl2 (} 1 mL). The mixture was stirred at room temperature for 30 hours. The reaction mixture was diluted with _CH2Cl2 , washed with water, saturated _NaHCO3 , brine _and dried over _{anhydrous Na2SO4} . Concentration in vacuo followed by silica gel flash chromatography ( _{CH2Cl2 -} MeOH- _NH4OH : 98:2:0.2 to 98:5:0.5) afforded compound 139 as an oil. Analytical HPLC retention time = 1.976 min (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 mL/min, monitored at 254 nm) and LC/MS M ⁺ +1 = ⁴⁴⁵⁺ .

Example 140

N-(3-chloro-4-fluorophenyl)-5-[[4-(propylamino)-1-piperidinyl]methyl]-pyrrolo[2,1-f][1,2,4 ] Triazin-4-amine

Compound 140 was prepared from Compound 1 in a similar manner to Compound 139. Compound 140 was a solid with an analytical HPLC retention time = 1.689 minutes (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 minutes, 4 mL/min, monitored at 254 nm) and LC/ MS M ⁺⁺ 1=417 ⁺ .

Example 141

1-[[4-[(3-Chloro-4-fluorophenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl]methyl]-4-piper Pyridyl alcohol

Compound 141 was prepared from 1E of Example 1 in a similar manner to Compound 1. Compound 141 was a solid with an analytical HPLC retention time = 1.803 min (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 mL/min, monitored at 254 nm) and LC /MS M ⁺⁺ 1=376 ⁺ .

Example 142

trans-4-[4-(3-chloro-4-fluoro-phenylamino)-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethyl]-cyclohexyl alcohol

A. Preparation of 4-chloro-pyrrolo[2,1-f][1,2,4]triazine-5-carbaldehyde

A solution of 4-chloro-5-methyl-pyrrolo[2,1-f][1,2,4]triazine (1.68 g, 10 mmole) in _CCl was bubbled with N for ₂₀ min, then NBS (3.74 g, 21 mmole) was added followed by dibenzoyl peroxide (242 mg, 1 mmole). The reaction mixture was placed in a 100°C oil bath and refluxed for 3 hours. After cooling to room temperature, it was filtered and the solvent was removed. The residue was suspended in _CH3OH (100 mL) and solid _NaHCO3 (5 g) was added. The reaction mixture was stirred vigorously for 1 hour, filtered and the solvent was removed. The residual oil was resuspended in DCM, filtered and concentrated to give crude dimethyl acetal which was treated with DCM (20 mL)/ _H2O (20 mL)/TFA (1 mL). After stirring vigorously for 1.5 h, it was neutralized with saturated aqueous NaHCO ₃ and extracted with DCM. The combined extracts were dried ( _Na2SO4 ), concentrated and chromatographed (3 x 15 cm silica gel column eluting with DCM) to afford the title compound as a solid (1.02 _g , 56%). MS: 182(M+H) ⁺ ; HPLC retention time: 0.79 minutes (Xterra 3.0×50 mm S7 column, 2-minute gradient, 5 ml/min);

B. [4-(tert-butyl-dimethyl-silyloxy)-cyclohexyl]-(4-chloro-pyrrolo[2,1-f][1,2,4]tri

Preparation of oxazin-5-yl)-methanol

A solution of trans-4-tert-butyl-dimethylsilyloxy-cyclohexylmagnesium bromide (Bioorg.and Med.Chem., 1996,6,201) in THF (4 equivalents) was slowly Add 4-chloro-pyrrolo[2,1-f][1,2,4]triazin-5-yl)-carbaldehyde (1.05 g 5.8 mmole) in THF (15 ml) to ice-cooled in solution. After 1 h, saturated aqueous _NH4Cl (15 mL) was added, and the aqueous layer was extracted with EtOAc/hexane (1:1) (50 mL x 2). _The combined organic extracts were dried over _Na2SO4 and concentrated in vacuo. The crude material was purified by radial chromatography (4 mm silica plate, gradient elution with 0-15% EtOAc in DCM) to afford the title compound: 189 mg of the cis-isomer, 496 mg of the trans-isomer and 415 mg of the mixture (total yield 48%, cis:trans ratio about 1:4). Cis-isomer: MS: 396 (M+H) ⁺ ; HPLC retention time: 2.10 min (Xterra 3.0×50 mm S7 column, 2 min gradient, 5 ml/min); trans-isomer: MS : 396 (M+H) ⁺ ; HPLC retention time 2.08 min (Xterra 3.0×50 mm S7 column, 2 min gradient, 5 ml/min).

C.[4-(3-Chloro-4-fluoro-phenylamino)-pyrrolo[2,1-f][1,2,4]triazin-5-yl]-[4-(1-methyl Preparation of -1-trimethylsilyl-ethoxy)-cyclohexyl]-methanol

Trans-[4-(tert-butyldimethylsilyloxy)-cyclohexyl]-(4-chloro-pyrrolo[2,1-f][1,2,4]triazine-5 A mixture of -yl)-methanol (840 mg, 2.13 mmole), 3-chloro-4-fluoroaniline (309 mg, 2.13 mmole) and NaHCO ₃ (536 mg, 6.39 mmole) in CH ₃ CN (10 mL) at 70° C. Heat overnight. The solvent was removed, and the residue was suspended in DCM, washed with water, and _dried over _Na2SO4 . Removal of solvent followed by radial chromatography (4 mm silica plate, eluting with a gradient of 0-2% _NH3 in MeOH (2N) in DCM) afforded the title compound as a solid (612 mg, 57%): MS: 506 (M+H) ⁺ ; HPLC retention time: 2.29 min (Xterra 3.0×50 mm S7 column, 2 min gradient, 5 ml/min).

D. trans-4-[4-(3-chloro-4-fluoro-phenylamino)-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethyl]- Preparation of hexanol

[4-(3-Chloro-4-fluoro-phenylamino)-pyrrolo[2,1-f][1,2,4]triazin-5-yl]-[4-(1-methyl A mixture of 1-trimethylsilyl-ethoxy)-cyclohexyl]-methanol (448 mg, 0.887 mmole), triethylsilane (1.03 g, 8.87 mmole) in TFA (8 ml) was resistant to Heat overnight at 75°C under _N2 in a press bottle. _The solvent was removed and the residue was dissolved in _CH3OH (10 mL) and solid _Na2CO3 (2.0 g) was added. After stirring vigorously for 1 h, the solvent was removed and the residue was partitioned between DCM (200 mL) and _H2O (50 mL). The organic phase was separated, dried over _Na2SO4 , and _the solvent was removed. Purification by radial chromatography (2 mm silica gel plates, eluting with a gradient of 0-4% _NH3 in MeOH (2N) in DCM) afforded the title compound (209 mg, 63%) as a solid: MS: 375 (M+H) ⁺ ; HPLC retention time: 1.49 min (Xterra 3.0 x 50 mm S7 column, 2 min gradient, 5 ml/min).

Example 143

cis-4-[4-(3-chloro-4-fluoro-phenylamino)-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethyl]-cyclohexyl alcohol

Likewise, the title compound is derived from cis-{4-(tert-butyldimethylsilyloxy)-cyclohexyl]-(4-chloro-pyrrolo[2,1-f][1,2, 4] Triazin-5-yl)-methanol Preparation: 375 (M+H) ⁺ ; HPLC retention time: 1.56 min (Xterra 3.0×50 mm S7 column, 2 min gradient, 5 ml/min).

Example 144

4-[4-(3-Chloro-4-fluoro-phenylamino)-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethyl]-cyclohexanone

cis-[4-(tert-butyldimethylsilyloxy)-cyclohexyl]-(4-chloro-pyrrolo[2,1-f][1,2,4]triazine-5 -yl)-methanol: (53 mg, 0.14 mmole), 4-methylmorpholine N-oxide (25 mg, 0.21 mmole), TPAP (5 mg, 0.1 equiv) and powdered 4A molecular sieves (100 mg) in DCM (3 mL) was stirred at RT under _N2 . After 5 hours, the solution was filtered and the solvent was removed. Radial chromatography (1 mm silica plate, gradient eluting with 0-5% _NH3 in MeOH (2N) in DCM) gave the title compound (25 mg, 47%) as a solid: MS: 373 ( M+H) ⁺ ; HPLC retention time: 1.50 min (Xterra 3.0 x 50 mm S7 column, 2 min gradient, 5 ml/min).

Example 145

4-amino-1-[4-(3-chloro-4-fluoro-phenylamino)-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethyl]-ring Hexanol

Powdered 3A molecular sieves (24 mg), (10 equiv) NH ₄ OAc (48 mg, 0.06 mmole) and NaCNBH ₃ (4 mg, 0.06 mmole) were added to 4-[4-(3-chloro-4-fluoro-phenylamino )-pyrrolo[2,1-f][1,2,4]triazin-5-ylmethyl]-4-hydroxy-cyclohexanone (24 mg, 0.06 mmole) in anhydrous MeOH (0.5 mL) solution; the reaction was stirred under nitrogen for 12 hours. The reaction mixture was filtered and 15% NaOH solution was added. After 10 minutes, the mixture was diluted with DCM (50 mL) and washed with water. The organic phase was dried (Na ₂ SO ₄ ) and the solvent was removed. The material was purified and isolated by preparative HPLC to give the title compound (3.5 mg, 15%) and the cis isomer (7.9 mg, 32%). Title compound: MS: 390 (M+H) ⁺ ; HPLC retention time: 2.070 min (XTERRA 4.6 x 50 mm S5 column, 3 min gradient, 4 ml/min). Cis isomer: MS: 390 (M+H) ⁺ ; HPLC retention time: 2.190 min (XTERRA 4.6×50 mm S5 column, 3 min gradient, 4 ml/min).

Example 146

(3R,4R)-4-amino-1-[[4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4triazin-5-yl] Methyl]piperidin-3-ol

Preparation of Compounds 146A and 146B:

(3R,4R)-4-Azido-3-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (146A):

(3S,4S)-4-Azido-3-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (146B)

Compounds 146A and 146B were obtained from compound 135L by optical resolution using normal-phase chiral preparative HPLC (Chiralpak AD) using MeOH-EtOH (50:50) as mobile phase. Compound 146A (first eluate) and compound 146B (second eluate) were obtained as single enantiomers with >99% ee. The absolute stereochemistry (3R, 4R) of compound 146A was determined by single X-ray crystallography.

Preparation of Compound 146C:

Trifluoroacetic acid (10 mL) was added to a solution _of compound 146A (1.76 g, 7.26 mmol) in anhydrous _CH2Cl2 (15 mL) cooled at 0 °C. The reaction mixture was stirred at 0 °C for 15 minutes, then allowed to warm to room temperature and stirred for 3 hours. The solvent and TFA were removed under reduced pressure, and the residue was azeotroped several _times with _CH2Cl2 to afford compound 146C as the TFA salt.

Preparation of Compound 146D:

Compound 146D was prepared as a TFA salt using compound 146B in a similar manner to compound 146C.

Preparation of Compound 146E:

Compound 146E was prepared by a method similar to Example 105 (Using Method 1), substituting compound 146C for piperidin-4-yl-carbamic acid tert-butyl ester. Compound 146E was a solid with an analytical HPLC retention time = 2.019 minutes (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% aqueous methanol containing 0.1% TFA over 4 minutes, 4 mL/min, monitored at 254 nm) and LC/ MS M ⁺⁺ 1=395 ⁺ .

Compound 146 was prepared from compound 146E by a method similar to compound 135. Compound 146 was a solid with analytical HPLC retention time = 1.213 min (ChromolithSpeedROD column 4.6 x 50 mm, 10-90% aqueous methanol containing 0.1% TFA over 4 min, 4 mL/min, monitored at 254 nm) and LC/MS M ⁺ +1 = 369 ⁺ . The enantiomeric excess (ee) of compound 146 was >99% (Chiralpak AD, 250 x 4.6 mm 10 μm, EtOH-MeOH- _Et2NH ; 50:50:0.1).

Alternative Preparation of Compound 146

Preparation of Compound 146F:

Solid _K2CO3 (235 g) was added to a solution of 1-methyl- _1,2,3,6 -tetrahydropiperidine HCl salt (227 g) in 570 mL of water, and the mixture was stirred at room temperature 30 minutes. The mixture was extracted with toluene (500 mL×3), and the combined extracts were dried over anhydrous MgSO ₄ . Filter to remove _MgSO4 , and place the filtrate in a 3-liter three-neck RB flask. K ₂ CO ₃ (22.7 g) was added to the filtrate, and the mixture was heated to gentle reflux (bath temperature 110° C.). Ethyl chloroformate (318 mL) was added slowly via an additional funnel over 2.5 hours (the reaction is very exothermic so slow addition and magnetic stirring is strongly recommended). Once the addition was complete, the mixture was refluxed for an additional 2.0 hours and cooled to room temperature. The reaction mixture was washed with water, brine, and dried over anhydrous MgSO ₄ . Filtration and concentration in vacuo afforded 188.6 g (72%) of compound 146F as an oil. ¹ H-NMR (400 MHz, CDCl ₃ ).

Preparation of compound 146G (racemic):

Solid m-CPBA (386 g, 1.72 mol, 77% max) was added in small portions to a solution of compound 146F (178.2 g, 1.15 mol) in 2 liters of anhydrous _CH2Cl2 at ₀ °C . The reaction mixture was stirred at 0°C for 1.0 hour and then at room temperature overnight. The precipitate was removed by filtration and _the filter cake was washed with _CH2Cl2 . The combined filtrate and washings were washed with 20% Na ₂ S ₂ O ₃ (3 L×3), saturated NaHCO ₃ (3 L×3), and dried over anhydrous Na ₂ SO ₄ . Filtration followed by concentration in vacuo afforded 170 g of compound 146G as an oil. This material was used directly in the next reaction step without further purification.

Preparation of compound 146H (chiral):

A mixture of compound 146G (140 g, 0.818 mol), NaN ₃ (68.9 g, 1.06 mol) and NH ₄ Cl (56.7 g, 1.06 mol) in ethanol (600 mL) and water (150 mL) was heated at 70 °C overnight. Once cooled to room temperature, the solids were removed by filtration and rinsed with ethanol. The combined filtrates were concentrated in vacuo to a small volume (ca. 80 mL), then diluted with water (500 mL) and extracted with EtOAc (500 mL x 4). The combined extracts were dried over _{anhydrous Na2SO4} . Filtration followed by concentration in vacuo and purification by flash chromatography on silica gel (hexane-EtOAc 7:3 to 6:4) gave the following fractions of compound 146G as an oil: 80.7 g of the first fraction (AP: > 98%), 22.7 grams of the second fraction (AP: 92-95%) and 15.8 grams of the third fraction (AP: <60%). This material was used directly in the next reaction step without further purification. The first and second fractions were combined and optically resolved using chiral preparative HPLC with the following conditions: Chiralpak AD column, eluting with MeOH-EtOH (1:1). The first eluting peak (Rt = 5.605 min) was collected to yield 47.52 g of compound 146H with >99% ee.

Preparation of compound 146I (chiral):

A solution of KOH (112 g, 1.7 mol, 85%) in 240 mL of water was added to a solution of compound 146H (36.42 g, 0.17 mol) in 480 mL of EtOH. The mixture was heated to reflux for 9.0 hours and the progress of the reaction was monitored by TLC. Once cooled to room temperature, the mixture was concentrated in vacuo to give a syrup. Solid NaCl was added and the mixture was extracted with EtOAc (500 mL x 3). The combined organic layers were dried over _{anhydrous Na2SO4} . Filtration followed by solvent removal under reduced pressure afforded 23 g (77%) of crude compound 146I as a solid. Trituration with diethyl ether (250 mL) gave 18.53 g of compound 146I as a solid (AP: 99%). The mother liquor was concentrated in vacuo, solid NaCl was added and further extracted with more EtOAc (250 mL x 4) to give an additional 4.2 g (AP: <85%) of crude compound 146I.

Compound 146 was prepared from Compound 146I following the procedure for Compound 146E.

Example 147

(3S,4S)-4-amino-1-[[4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl ]methyl]3-piperidin-3-ol

Preparation of Compound 147A:

Compound 147A was prepared by a method similar to Example 105 (using method 1), substituting compound 130D for tert-butyl piperidin-4-yl-carbamate. Compound 147A was a solid with an analytical HPLC retention time = ? min (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 ml/min, monitored at 254 nm) and LC/MS M ⁺ +1 = 395 ⁺ .

Compound 147 was prepared from Compound 147A by a method similar to Compound 135. Compound 147 was a solid with an analytical HPLC retention time = 1.213 minutes (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% aqueous methanol containing 0.1% TFA over 4 minutes, 4 ml/min, monitored at 254 nm) and LC/ MS M ⁺⁺ 1=369 ⁺ . The enantiomeric excess (ee) of compound 147 was >99% (Chiralpak AD, 250 x 4.6 mm 10 microns, EtOH-MeOH- _Et2NH : 50:50:0.1).

Example 148

(3R,4R)-4-amino-1-[[4-[(3-methoxy-4-fluorophenyl)amino]pyrrolo[2,1-f][1,2,4]triazine -5-yl]methyl]piperidin-3-ol

Compound 148 was prepared from compound 146C by a method similar to compound 135. Compound 148 was a solid with analytical HPLC retention time = 1.187 minutes (Chromolith SpeedROD column 4.6 × 50 mm, 10-90% methanol in water, containing 0.1% TFA, experienced 4 minutes, 4 ml/min, monitored at 254nm) and LC/ MS M ⁺⁺ 1=387 ⁺ . The enantiomeric excess (ee) of compound 148 was >99% (Chiralpak AD, 250 x 4.6 mm 10 μm, EtOH-MeOH- _Et2NH : 50:50:0.1).

Example 149

(3S,4S)-4-amino-1-[[4-[(3-methoxy-4-fluorophenyl)amino]pyrrolo[2,1-f][1,2,4]triazine -5-yl]methyl]-piperidin-3-ol

Compound 149 was prepared from compound 146D by a method similar to compound 135. Compound 149 was a solid with analytical HPLC retention time = 1.187 min (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% aqueous methanol containing 0.1% TFA over 4 min, 4 ml/min, monitored at 254 nm) and LC /MS M ⁺⁺ 1=387 ⁺ . The enantiomeric excess (ee) of compound 149 was >99% (Chiralpak AD, 250 x 4.6 mm 10 μm, EtOH-MeOH- _Et2NH : 50:50:0.1).

Example 150-200

Compounds 150-200 (with HPLC annotation (b)) were prepared in the same way as compound 146 from 146I.

^a Gradient time of 2 minutes for HPLC. ^b Gradient time of 2 min for HPLC (Phenom-prime S5 C18 4.6 x 30 mm column).

Example 201

rac-(3S,4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazine-5 -yl}methyl)piperidin-3-ol

Preparation of 201A4-azido-3-oxopiperidine-1-carboxylic acid (±)-tert-butyl ester

Anhydrous DMSO (0.28 mL, 3.79 mmole) was added to a stirred solution of oxalyl chloride (0.172 mL, 1.96 _mmol ) in 6 mL of anhydrous _CH2Cl2 at -78 °C under argon. After 10 min, a solution of compound 135L (396 mg, 1.63 mmol) in 4.5 mL of anhydrous _{CH2Cl2 was} added dropwise, and the reaction mixture was stirred at -78 °C for 30 min. Triethylamine (1.38 mL, 10.0 mmol) was added and the reaction mixture was allowed to warm to room temperature. 2.0 mL _of pH 7.0 buffer was added and the mixture was extracted with _CH2Cl2 (x3). The combined organic layers were washed with brine and dried over _{anhydrous Na2SO4} . Concentration in vacuo gave crude 201A as an oil, which was used directly in the next reaction step. ¹ H-NMR (400MHz, CDCl ₃ ): 4.30 (d, J=17.84, 1H), 4.05 (m, 1H), 3.90-4.00 (m 1H), 3.45 (m, 1H), 2.85 (m, 1H) , 2.33(m, 1H), 1.86(m, 1H), 1.47(s, 9H).

201B. Preparation of 4-azido-3-hydroxypiperidine-1-carboxylic acid (±)-tert-butyl ester

L-Selectride (1.0 M in THF, 0.98 mL, 0.98 mmol) was added to a solution of compound 201A prepared above in anhydrous THF (2 mL) cooled at -78 °C. The mixture was stirred at -78°C for 2.0 hours. Sat. _NH4Cl (2 mL) was added, and the reaction mixture was allowed to warm to room temperature. The mixture was diluted with water and extracted with EtOAc (3x). The combined organic layers were washed once with brine and dried over _{anhydrous Na2SO4} . Concentration in vacuo followed by silica gel flash chromatography (Hexane-EtOAc 4:1) gave 44 mg of compound 201B as an oil.

¹ H-NMR (400mHz, CDCl ₃ ): 3.84(m, 1H), 3.69(m, 1H), 3.58(m, 2H), 3.40(m, 1H), 3.30(m, 1H), 1.96(m, 1H), 1.73(m, 1H), 1.46(s, 9H). 201C. Preparation of (±)-4-azidopiperidin-3-ol

Compound 201B (44 mg, 0.18 mmol) was treated with a mixture of _CH2Cl2 and TFA (1:1, 2 mL) for 30 _min . The volatiles were removed under reduced pressure, and the residue _was azeotroped three times with heptane- _CH2Cl2 to give compound 201C as TFA salt, which was directly used in the next reaction without step of further purification.

Preparation of 201D:

Compound 201D was prepared as a solid from compound 201C by a method similar to compound 146E. It had an analytical HPLC retention time = 1.795 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 mL/min, monitored at 220 nm) and LC/MS M ⁺ = 395 .

Compound 201 was prepared from Compound 201D in a similar manner to Compound 146. It had an analytical HPLC retention time = 1.169 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ = 369 .

Examples 202A and 202B

(3S, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidin-3-ol (enantiomer A, chiral)

and

(3R, 4S)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidin-3-ol (enantiomer B, chiral)

Compound 202A (15 mg) and compound 202B (15 mg) were obtained by optical resolution of compound 201 (30 mg) using the following method: Hexane-isopropanol-diethylamine (50: 50:0.1) elution and detection at 220nm Chiralpak AD chiral preparative column. The first elution peak corresponds to compound 202A with ee%≥98% (retention time=4.337 minutes); the second elution peak corresponds to compound 202B with ee%≥98% (retention time=6.050 minutes).

Example 203

(3S, 4R)-4-amino-1-({4-[(3-methylphenyl)amino]pyrrolo[2,1-F][1,2,4]triazin-5-yl} Methyl)piperidin-3-ol (chiral)

Preparation of compounds 203A and 203B (chiral):

and

196.5 mL of 48% HBr was added dropwise to a -60°C cooled solution of compound 146G (100 g, 0.585 mol) in 2 liters of chloroform via an addition funnel while keeping the internal temperature below -60°C. Once the addition was complete, the reaction mixture was stirred for an additional 1.0 hour at -60°C. The reaction mixture was allowed to warm to room temperature and washed with water (1 L x 2), brine (1 L), and dried over anhydrous MgSO ₄ . Filtration followed by concentration in vacuo afforded 134.2 g (91%) of the crude compound as an oil (racemic mixture 203A and 203B).

¹ H-NMR (400MHz, CDCl ₃ ): 4.25 (m, 1H), 4.15 (q, J=7.10, 2H), 4.00 (m, 1H), 3.90 (bs, 1H), 3.75 (m, 1H), 2.85-3.15(m, 2H), 2.32(m, 1H), 2.00(m, 1H), 1.28(t, J=7.10, 3H).

Compounds 203A and 203B were obtained from the optical resolution of the above racemic mixtures by normal phase chiral preparative HPLC (Chirlapak AD) using _CH3CN as mobile phase. 54.77 g of compound 203B (first eluent, Rt = 5.861 min) and 53.71 g of compound 203A (second eluent, Rt = 8.719 min) as a single enantiomer with >99% ee get. The absolute stereochemistry (3R, 4S) of compound 203B was determined from single X-ray crystallographic analysis of compound 203.

Preparation of compound 203C (chiral):

Imidazole (21.8 g, 0.32 mol), followed by tert-butyldimethylsilyl chloride (38.5 g, 0.258 mol) was added to compound 203A (53.7 g, 0.213 mol) in 250 mL of DMF at 0 °C in the solution. The reaction mixture was stirred overnight at ambient temperature. Diethyl ether (1 L) followed by water (1 L) was added to the reaction mixture at 0°C. Separate the organic layer. The aqueous layer was extracted with ether (1 L×2), and the combined organic layers were washed with 10% LiCl (750 mL×3), dried over anhydrous MgSO ₄ . Filtration followed by concentration in vacuo gave crude compound 203C as an oil which was used without further purification.

Preparation of compound 203D (chiral):

Compound 203D was prepared from Compound 203B by a method similar to Compound 203C.

Preparation of compound 203E (chiral):

NaN ₃ (15.3 g, 0.234 mol) was added to a solution of compound 203C (0.213 mol) in 300 mL of DMSO, and the mixture was heated at 85° C. for 12 hours. Additional _NaN3 (15.0 g, 0.230 mol) was added and the reaction mixture was heated overnight. Once cooled to room temperature, ice water was added to the reaction mixture and extracted with diethyl ether (1 L x 3). The combined organic layers were washed once with brine (1 L) and dried over anhydrous _MgSO4 . Filtration followed by concentration in vacuo gave crude compound 203 E as an oil which was used without further purification.

Preparation of compound 203F (chiral):

Compound 203F was prepared from Compound 203D by a method similar to Compound 203E.

Preparation of compound 203G (chiral):

A mixture of compound 203E (0.213 mol) prepared above and TBAF xH ₂ O (67 g, 0.255 mol) in 200 mL THF was stirred at room temperature for 3.0 hours. Diethyl ether (1 L) was added and the mixture was washed with water (1L). The aqueous phase was extracted with ether (1 L x 2). The combined organic layers were washed once with water (1 L) and dried over anhydrous _MgSO4 . Concentration in vacuo followed by silica gel flash chromatography ( _CH2Cl2 -EtOAc: 4:1) gave 29.8 g of compound 203G as _an oil. A second flash chromatography on silica gel (Hexane-EtOAc: 6.5:3.5) afforded 20 g (44%) of compound 203G as an oil.

¹ H-NMR (400 MHz, CDCl ₃ ): 4.15 (q, J=7.10, 2H), 3.86 (bs, 1H), 3.70 (bs, 1H), 3.65 (m, 1H), 3.47 (dd, J=3.20 , J=13.62, 1H), 3.35(m, 1H), 2.02(m, 1H), 1.79(bs, 1H), 1.28(t, J=7.10, 3H).

Preparation of compound 203H (chiral):

Compound 203H was prepared from Compound 203F by a method similar to Compound 203G.

Preparation of compound 203I (chiral):

Compound 203I was prepared from compound 203G by a method similar to compound 146I. Compound 203I is a solid with > 99 ee%.

¹ H-NMR (400MHz, CDCl ₃ ): 3.80(m, 1H), 3.44(m, 1H), 3.04(m, 2H), 2.72(m, 1H), 2.69(m, 1H), 1.90(m, 1H), 1.75(m, 1H).

Preparation of compound 203J (chiral):

Compound 203J was prepared from compound 203H by a method similar to compound 146I. Compound 203J is a solid with > 99 ee%.

Compound 203 was prepared from compound 1B, m-methylaniline and compound 203I by a similar method to compound 146. It has analytical HPLC retention time = 1.278 min (ChromolithSpeedROD 4.6 x 50 mm column, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 mL/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1 =353.

Examples 204-211

Compounds 204-211 (with HPLC annotation (b)) were prepared similarly from Compound IB, the corresponding aniline and Compound 203I as used for Compound 146.

Examples 212-219

Compounds 212-219 (with HPLC annotation (b)) were prepared similarly from Compound IB, the corresponding aniline and Compound 203J as used for Compound 146.

Example 220

(3R,4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]-triazine-5- Base}methyl)-piperidin-3-ylcarbamate

Preparation of compound 220A (chiral):

Trichloroacetylisocyanate (0.075 mL, 0.6 mmol) was added to a solution of compound 146A (121 mg, 0.5 _mmol ) in 1 mL of anhydrous _CH2Cl2 cooled at 0 °C. The reaction mixture was stirred at 0 °C for 1.0 h. MeOH (0.5 mL) was added and the reaction mixture was concentrated in vacuo to afford crude compound 220A as a foam.

¹ H-NMR (400MHz, CDCl ₃ ): 8.35(bs, 1H), 4.67(bs, 1H), 3.90(m, 1H), 3.65(m, 2H), 3.25(m, 2H), 2.02(m, 1H), 1.60(m, 1H), 1.38(s, 9H).

Preparation of compound 220B (chiral):

A solution of 20% _{aqueous K2CO3} (2 mL) was added to a solution of 220A (0.5 mmol) in 3 mL of anhydrous MeOH, and the reaction mixture was stirred at room temperature for 2.0 h. Water (15 mL) was added and MeOH was removed by rotary evaporation. The mixture was extracted with EtOAc (x2) and dried over _{anhydrous Na2SO4} . Concentration in vacuo gave crude 220B as an oil. ¹ H-NMR (400MHz, CDCl ₃ ): 4.70 (bs, 2H), 4.50 (m, 1H), 3.90 (bs, 1H), 3.68 (m, 1H), 3.50 (m, 1H), 3.03 (m, 1H), 1.90(m, 1H), 1.50(m, 1H), 1.38(s, 9H).

Preparation of compound 220C (chiral):

A mixture _of 220B (0.5 mmol) in 5 mL anhydrous _CH2Cl2 and 5 mL TFA was stirred at 0 °C for 1.0 h. The mixture was concentrated in vacuo, azeotroped several times with _{CH2Cl2 -} MeOH-hexane, and dried under high vacuum to give crude 220C as an oil.

Preparation of compound 220D (chiral):

Compound 220D was prepared from compound 220C in a similar manner to compound 146E. It had an analytical HPLC retention time = 1.793 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 mL/min, monitored at 220 nm) and LC/MS M ⁺ = 438 .

Compound 220 was prepared from compound 220D in a similar manner to compound 146. It had an analytical HPLC retention time = 1.310 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ = 412 .

Examples 221-227

Compounds 221-227 (with HPLC annotation (b)) were prepared analogously as Compound 146 from Compound IB, the corresponding aniline and Compound 220C.

Example 229

(3R, 4S)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidin-3-ylcarbamate

229A. Preparation of Compound 229A (Chiral)

Compound 229A was prepared from compound 203I by a method similar to step 2 of compound 135E.

229B. Preparation of compound 229B (chiral)

Compound 229B was prepared from Compound 229A by a method similar to Compound 220A.

229C. Preparation of compound 229C (chiral)

Compound 229C was prepared from Compound 229B by a method similar to Compound 220B.

229D. Preparation of Compound 229D

Compound 229D was prepared from Compound 229C by a method similar to Compound 220C.

Compound 229 was prepared from compound 229D by a similar method to compound 146. It had an analytical HPLC retention time = 1.229 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ = 412 .

Examples 230-236

Compounds 230-236 (with HPLC annotation (b)) were prepared analogously as compound 146 from compound IB, the corresponding aniline and compound 229D.

Example 237

(3R, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)-3-methylpiperidin-3-ol (chiral, enantiomer A)

Preparation of compound 237A (racemic):

Solid _NaHCO3 (4.2 g, 49 mmol) followed by methyl chloroformate (9.3 mL, 120 mmol) was added dropwise via syringe to a solution of N-benzyltetrahydropyridine (100 mmol) in 150 mL of benzene at room temperature . The reaction mixture was heated to reflux for 3 hours. After cooling to room temperature, the volatiles were removed by evaporation under reduced pressure, and the residue was dissolved in EtOAc (100 mL) and washed with water (20 mL×2), 0.5M HCl (20 mL) and brine (20 mL) and dried over anhydrous MgSO ₄ . The reaction mixture was concentrated in vacuo and residual benzyl chloride was further removed under high vacuum (bp. 42-50°C/2 mmHg, bath temperature: 75-80°C) to give compound 237A as a viscous syrup. Silica gel flash chromatography (hexane-EtOAc: 9.5:0.5 to 9:1) afforded 11.77 g (yield: 76%) of compound 237A as an oil.

¹ H-NMR (400MHz, CDCl ₃ ): 5.52 (bs, 1H), 3.78 (m, 2H), 3.70 (s, 3H), 3.46 (m, 2H), 2.08 (m, 2H), 1.68 (s, 3H).

Compound 237B (racemic) Preparation:

A mixture _of compound 237A (775 mg, 5.0 mmol) and m-CPBA (1.21 g, 7.0 mmol, 77% max) in 10 mL of anhydrous _CH2Cl2 was stirred overnight at room temperature. The precipitate was removed by filtration, and the filtrate was washed with 10% Na ₂ S ₂ O ₃ , saturated NaHCO ₃ and brine, and dried over anhydrous MgSO ₄ . Concentration in vacuo afforded crude compound 237B as an oil.

¹ H-NMR (400 MHz, CDCl ₃ ): 3.65-3.75 (m, 2H), 3.68 (s, 3H), 3.60 (m, 1H), 3.33 (m, 2H), 3.12 (m, 1H), 2.07 ( m, 1H), 1.92(m, 1H), 1.35(s, 3H).

Compound 237C (racemic) was prepared:

NaN ₃ (810 mg, 12.4 mmol) was added to a solution of compound 237B (5.0 mmol) in 10 mL DMF in 10 mL of acetone-H ₂ O 2:1 mixture, and the mixture was heated at 80° C. overnight . After cooling to room temperature, EtOAc was added and the mixture was washed with water, 10% aqueous LiCl and dried over _{anhydrous Na2SO4} . Concentration in vacuo gave 1.09 g of compound 237C as an oil.

¹ H-NMR (400MHz, CDCl ₃ ): 3.70(s, 3H), 3.72(m, 1H), 3.50(m, 2H), 3.35(m, 1H), 3.20(bs, 1H), 2.03(m, 1H), 1.63(m, 1H), 1.20(s, 3H).

Compound 237D (racemic) Preparation:

A mixture of compound 23 7C (428 mg, 2.0 mmol) and 150 mg of Pd(OH) ₂ in 10 mL of MeOH was stirred under hydrogen (cylinder) for 4.0 h. The catalyst was removed by filtration and the filtrate was concentrated in vacuo to give a crude residue.

The residue was dissolved in 5 mL of _{anhydrous CH2Cl2} , to which _Boc2O (460 mg, 2.10 mmol) and triethylamine (0.334 mL, 2.40 mmol) were added. After stirring at room temperature overnight, the reaction mixture _was diluted with _CH2Cl2 and washed with 10% citric acid, sat. _NaHCO3 and _dried ( _Na2SO4 ). Concentration in vacuo followed by silica gel flash chromatography (Hexane-EtOAc: 7:3 to 1:1) gave 330 mg of 237D as an oil.

¹ H-NMR (400MHz, CDCl ₃ ): 4.78(m, 1H), 3.90-4.40(m, 3H), 3.70(s, 3H), 3.60(m, 1H), 2.70-3.00(m, 2H), 1.80 (m, 1H), 1.45 (s, 9H), 1.40 (m, 1H), 1.10 (s, 3H). Compound 237E (racemic) Preparation:

Compound 237E was prepared from compound 237D by a method similar to compound 146I.

Compound 237F (racemic) Preparation:

Compound 237F was prepared from Compound 237E by a method similar to Compound 1D.

Compound 237G (racemic) Preparation:

Compound 237G was prepared from compound 237F by a method similar to compound 1E. It has analytical HPLC retention time = 1.262 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 mL/min, monitored at 220 nm) and LC/MS M ⁺ +1 =383.

Compound 237 was separated by chiral preparative HPLC (Chiralpak AD, 250 x 4.6 mm, 10 μm, eluting with EtOH/MeOH/DEA 50:50:0.1) from 237G with ee%≥99% of the first peak (Rt = 5.390 min) was obtained. It has an analytical HPLC retention time = 1.51 min (Chromolith SpeedROD 4.6 x 50 mm column, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=383.

Example 238

(3S, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)-3-methylpiperidin-3-ol (chiral, enantiomer B)

Compound 238 was separated by chiral preparative HPLC (Chiralpak AD, 250 x 4.6 mm, 10 microns, eluting with EtOH/MeOH/DEA 50:50:0.1) from 237G to a second peak with ee%≥99% (Rt = 8.523 min) was obtained. It has an analytical HPLC retention time = 1.51 min (Chromolith SpeedROD 4.6 x 50 mm column, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=383.

Example 239

(3R, 4R, 5S)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazine-5 -yl}methyl)piperidine-3,5-diol

239A. Preparation of (±)-ethyl 3-acetoxy-4-bromopiperidine-1-carboxylate

A mixture of compound 203A/B (racemic mixture) (107 g, 371 mmol) and acetic anhydride (101 mL) in 165 mL of anhydrous pyridine was stirred at room temperature for 2.0 hours. The solvent was _removed under reduced pressure, and the residue was diluted with water and basified with _K2CO3 . The mixture was extracted with chloroform (250 mL×3), and the combined extracts were washed with water and dried over anhydrous MgSO ₄ . Concentration in vacuo afforded crude 239A as an oil.

¹ H-NMR (400 MHz, CDCl ₃ ): 4.90 (bs, 1H), 4.13 (q, J=7.08, 2H), 4.10 (m, 1H), 3.89 (m, 1H), 3.60 (m, 1H), 3.50(m, 2H), 2.34(m, 1H), 2.08(s 3H), 1.93(m, 1H), 1.27(t, J=7.80, 3H).

239B. Preparation of (±)-ethyl 5-acetoxy-5,6-dihydropyridine-1(2H)-carboxylate

A mixture of compound 239A (371 mmol) and DBU (92 g, 604 mmol) was heated at 90-110 °C for 30 minutes. After cooling to room temperature, the mixture was diluted with toluene and stirred for an additional 30 minutes. The precipitate was removed by filtration and rinsed with toluene. The combined filtrates were washed with 1.0N HCl, water, and dried over anhydrous _MgSO4 . Concentration in vacuo afforded 67.56 g (yield: 85.4%) of 239B as an oil.

¹ H-NMR (400MHz, CDCl ₃ ): 6.00 (bs, 1H), 5.90 (bs, 1H), 5.20 (m, 1H), 4.20 (q, J=7.08, 2H), 4.19 (m, 1H), 3.85(m, 1H), 3.80(m, 1H), 3.55(m, 1H), 2.08(s, 3H), 1.28(t, J=7.08, 3H).

239C. Preparation of (±)-ethyl 5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate

A solution of 0.2N NaOH in EtOH (65 mL) was added to a solution of compound 239B (10.5 g, 49.2 mmol) in 30 mL of EtOH, and the mixture was stirred at 0 °C for 30 min. After warming to room temperature, the reaction mixture was neutralized with glacial acetic acid. Concentration in vacuo gave 8.5 g of 239C as an oil.

¹ H-NMR (400 MHz, CDCl ₃ ): 5.90 (m, 1H), 5.82 (bs, 1H), 4.20 (bs, 1H), 4.18 (q, J=7.08, 2H), 4.04 (m, 1H), 3.85(m, 1H), 3.55(m, 2H), 1.28(t, J=7.08, 3H).

239D. Preparation of (±)-ethyl 5-(tert-butyldimethylsilyloxy)-5,6-dihydropyridine-1(2H)-carboxylate

Imidazole (9.5g, 140.0mmol) and tert-butyldimethylsilyl chloride (15.5g, 102.8mmol) were added to compound 239C (16g, 93.5mmol) in 150ml anhydrous CH ₂ Cl at 0°C ₂ , and the mixture was stirred overnight at ambient temperature. The reaction mixture was diluted with ether (500 mL) and water (1 L) at 0°C. The organic phase was separated, washed with 10% LiCl (150 mL x 3), dried over anhydrous MgSO ₄ . Concentration in vacuo followed by flash chromatography (hexane-EtOAc: 15:1 to 10:1) afforded 25.2 g (yield: 94.4%) of 239D as an oil.

239E. Preparation of (±)-ethyl 5-hydroxy-7-oxa-3-aza-bicyclo[4.1.0]heptane-3-carboxylate

Solid m-CPBA (39.6 g, 176.6 mmol, 77% max) was added in small portions to _a solution of compound 239D (25.2 g, 88.3 mmol) in 250 mL of _CH2Cl2 at 0 °C while allowing the internal temperature to Keep below 0°C during the addition. The mixture was stirred at 0°C for 30 minutes then at room temperature for 4.0 hours. More solid m-CPBA (39.6 g, 176.6 mmol, 77% max) was added in small portions and the mixture was stirred at room temperature for three days. The precipitate was removed by filtration, and the filtrate was washed with ₂₀ % _Na2S2O3 (500 mL×3), saturated _NaHCO3 ( ₅₀₀ mL×3) and brine (250 mL), dried over _{anhydrous Na2SO4} . Purification by silica gel flash chromatography (Hexane-EtOAc: 9.5:0.5 to 9:1) afforded 4.33 g of 239E (low Rf) as an oil.

239F. Preparation of (±)-ethyl 5-(tert-butyldimethylsilyloxy)-7-oxa-3-aza-bicyclo[4.1.0]heptane-3-carboxylate

Compound 239F was prepared from 239D by the same reaction as compound 239E as an oil.

239G. Preparation of (±)-ethyl 5-hydroxy-7-oxa-3-aza-bicyclo[4.1.0]heptane-3-carboxylate

A solution of TBAF (17.2 mL, 17.2 mmol) in THF was added to a solution of compound 239E (4.33 g, 14.4 mmol) in 10 mL of anhydrous THF. The mixture was stirred overnight at room temperature. Water was added and the reaction mixture was extracted with EtOAc (100 mL x 3). The combined organic _layers were washed with brine and dried over _Na2SO4 . Concentration in vacuo followed by silica gel flash chromatography (hexane-EtOAc: 1:1 to 1:2) gave 2.01 g (yield: 74.5%) of 239G as an oil.

¹ H-NMR (400MHz, CDCl ₃ ): 4.13(q, J=7.08, 2H), 4.05(m, 1H), 3.80(m, 1H), 3.70(d, J=10.15, 1H), 3.62(m , 1H), 3.45(m, 2H), 3.13(dd, J=10.15, J=7.63, 1H), 2.40 and 2.25(part, 1H), 1.27(t, J=7.08, 3H).

Preparation of 239H.(±)-ethyl 5-hydroxy-7-oxa-3-aza-bicyclo[4.1.0]heptane-3-carboxylate

A solution of TBAF (67.6 mL, 67.6 mmol) in THF was added to a solution of compound 239F (13.6 g, 45.1 mmol) in 10 mL of anhydrous THF. The mixture was stirred overnight at room temperature. Water was added and the reaction mixture was extracted with EtOAc (250 mL x 3). The combined organic layers were washed with brine and dried over _{anhydrous Na2SO4} . Concentration in vacuo followed by silica gel flash chromatography (hexane-EtOAc: 1:1 to 1:2) gave 6.03 g (yield: 75%) of 239H as an oil.

¹ H-NMR (400MHz, CDCl ₃ ): 4.15 (m, 1H), 4.13 (q, J=7.08, 2H), 3.35 (m, 1H), 3.75 (m, 1H), 3.52 (m, 1H), 3.32(m, 1H), 3.25(m, 2H), 3.10 and 2.55 (part, 1H), 1.27(t, J=7.08, 3H).

239I. Preparation of (±)-ethyl 4-azido-3,5-dihydroxypiperidine-1-carboxylate

NaN ₃ (3.5 g, 53.4 mmol) and NH ₄ Cl (2.3 g, 42.72 mmol) were added to a solution of compound 239G (2.0 g, 10.7 mmol) in 2-methoxyethanol (40 mL). The reaction mixture was heated at 125°C overnight. After cooling to room temperature, the solvent was removed under reduced pressure and the residue was dissolved in EtOAc (50 mL), washed with water (10 mL x 2), and dried over _{anhydrous Na2SO4} . Concentration in vacuo followed by silica gel flash chromatography (Hexane-EtOAc: 1:1 to 1:2) afforded 0.64 g of 239I (higher Rf) as a crystalline material. Stereochemistry was confirmed by single X-ray crystallography.

239J. Preparation of (±)-ethyl 4-azido-3,5-dihydroxypiperidine-1-carboxylate

239J was prepared from 239H by a similar reaction to compound 239I as a crystalline material.

239K. Preparation of (meso)-4-azido-3,5-bis(methoxymethoxy)piperidine

i-Pr2NEt (5.9 mL, 33.4 mmol), followed by MOMCl (1.69 mL, 22.2 mmol) was added to a solution of compound 239I (640 mg, 2.78 mmol) in anhydrous _{CH2Cl2 (} 5 mL). The reaction mixture was heated at 60 °C overnight. After cooling to room temperature, the reaction was diluted with _CH2Cl2 , washed with 10% citric acid, _{saturated NaHCO3} and brine, and dried over _{anhydrous Na2SO4} . Concentration in vacuo gave the crude intermediate as an oil, which was used directly in the next reaction step without further purification.

A mixture of the intermediate prepared above and KOH (1.84 g, 85%) in 8 mL of EtOH and 4 mL of water was heated to reflux overnight. After cooling to room temperature, the solvent was removed under reduced pressure and the residue was diluted with water and extracted with _CH2Cl2 ( _x2 ). The combined organic layers were dried over _{anhydrous Na2SO4} . Concentration in vacuo afforded 662 mg of 239K as an oil. This material was used directly in the next reaction step without further purification.

¹ H-NMR (400MHz, CDCl ₃ ): 4.73(q, J=6.82Hz, 4H), 3.41(s, 6H), 3.20-3.40(m 5H), 2.46(m, 2H).

Preparation of 239L(±)-(3R,5R)-rel-4-azido-3,5-bis(methoxymethoxy)-piperidine

Compound 239L was prepared from 239J by a similar reaction to compound 239K as a crystalline material.

¹ H-NMR (400MHz, CDCl ₃ ): 4.72(q, J=6.82 Hz, 4H), 3.92(m, 1H), 3.79(m, 1H), 3.49(m, 1H), 3.42(s, 3H) , 3.41(s, 1H), 3.25(m, 1H), 3.03(m, 1H), 2.70(m, 1H), 2.53(m, 1H).

239M (±)-(3S, 4R, 5R)-rel-4-azido-1-((4-(3-methoxyphenylamino)-pyrrolo[1,2-f][1, Preparation of 2,4]triazin-5-yl)methyl)-3,5-bis(methoxymethoxy)-piperidine

Compound 239M was prepared from 239K by the same reaction as compound 146E as a foam. Compound 239M has an analytical HPLC retention time = 2.582 minutes (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 minutes, 4 ml/min, monitored at 254nm) and LC/MS M ⁺ +1=502 ⁺ .

239N.(3S, 4R, 5R)-rel-4-azido-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4] Preparation of Triazin-5-yl)methyl)piperidine-3,5-diol

A mixture of compound 239M (0.65 mmol) and 6N HCI (2 mL) in 3 mL THF was heated at 50 °C for 2 hours. After cooling to room temperature, the mixture was basified with NaOH, extracted with EtOAc, and dried over _{anhydrous Na2SO4} . Concentration in vacuo afforded 260 mg of solid 239N. Compound 239N has an analytical HPLC retention time = 2.063 minutes (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 minutes, 4 ml/min, monitored at 254 nm) and LC/MS M ⁺ 1 = 411 ⁺ .

A mixture of compound 239N (260 mg, 0.633 mmol) and Ph3P ( ₃₃₂ mg, 1.27 mmol) in THF (3 mL) and water (0.3 mL) was heated at 70 °C overnight. After cooling to room temperature, the mixture was diluted with water, acidified with 2N HCl, and washed with EtOAc (2x). The aqueous layer was basified with 2N NaOH and extracted with EtOAc (2x). The combined organic layers were washed once with water and dried over _{anhydrous Na2SO4} . Concentration in vacuo followed by trituration with ether afforded 180 mg of 239 as a solid. Compound 239 has an analytical HPLC retention time = 1.211 minutes (Chromolith SpeedROD column 4.6 × 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 minutes, 4 ml/min, monitored at 254nm) and LC/MS M ⁺ +1=385 ⁺ .

Example 240

(3R/S, 5R/S)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazine -5-yl}methyl)piperidine-3,5-diol

Compound 240 was prepared from 239L by a method similar to compound 239. Compound 240 was a solid with an analytical HPLC retention time = 1.045 min (Chromolith SpeedROD column 4.6 x 50 mm, 10-90% aqueous methanol containing 0.1% TFA over 4 min, 4 mL/min, monitored at 254 nm) and LC/ MS M ⁺⁺ 1=384 ⁺ .

Example 241A

(3S,5S)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidine-3,5-diol (enantiomer A, chiral)

Compound 241A (enantiomer A) was prepared from 240 by chiral preparative HPLC separation (using Chiralpak AD and eluting with EtOH/DEA: 100/0.1) as the first peak with ee > 99% ( Rt = 5.827 minutes).

Compound 241A was a solid with an analytical HPLC retention time = 1.044 min (ChromolithSpeedROD column 4.6 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 mL/min, monitored at 254 nm) and LC/MS M ⁺ +1 = 384 ⁺ .

Example 241B

(3R,5R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidine-3,5-diol (enantiomer B, chiral)

Compound 241B (enantiomer B) was prepared from 240 by chiral preparative HPLC separation (using Chiralpak AD and eluting with EtOH/DEA: 100/0.1) as the second peak with ee > 96% ( Rt = 8.430 min).

Examples 242-246

Compounds 242-246 (with HPLC annotation (b)) were prepared from 239L by a method similar to compound 239.

^a Chiral normal phase HPLC conditions: Chiralpak AD, isocractic, eluting with EtOH-DEA: 100/0.1.

Example 247

rac-5-{[(3R,4R)-4-amino-3-methoxypiperidin-1-yl]methyl}-N-(3-methoxyphenyl)pyrrolo[2,1- f][1,2,4]triazin-4-amine

247

247A. Preparation of (3R,4R)-rel-tert-butyl 4-azido-3-methoxypiperidine-1-carboxylate

95% NaH (40.0 mg, 1.58 mmol) was added to a stirred mixture of 146A (320 mg, 1.32 mmol) and iodomethane (0.25 mL, 3.96 mmol) in 4 mL of anhydrous THF under nitrogen at room temperature. The mixture was stirred at room temperature for 15 hours and then quenched by adding 30 ml of water. The aqueous solution was extracted with EtOAc (2 x 40 mL). The combined EtOAc extracts were washed with brine (30 mL), dried ( _MgSO4 ), filtered and concentrated in vacuo to give 310 mg (Yield: 92%) of 247A. It has analytical HPLC retention time = 2.86 minutes (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% methanol in water, containing 0.1% TFA, over 4 minutes, 4 ml/min, monitored at 220 nm) and LC/ MS M ⁺⁺ Na=279.

247B.5-(((3R,4R)-rel-4-azido-3-methoxypiperidin-1-yl)methyl)-N-(3-methoxyphenyl)pyrrolo[ Preparation of 1,2-f][1,2,4]triazin-4-amine

TFA (2.00 mL, 26.0 _mmol ) was added to a stirred solution of 247A (310 mg, 1.21 mmol) in 2 mL _CH2Cl2 at room temperature. The mixture was stirred at room temperature for 15 minutes and concentrated in vacuo to give 390 mg of (3R,4R)-rel-4-azido-3-methoxypiperidine as its TFA salt. Compound 247B was prepared from this TFA salt (52.0 mg, 0.19 mmol) by a method similar to that described for 146E. It has analytical HPLC retention time = 1.97 min (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=409.

247C.5-(((3R,4R)-4-amino-3-methoxypiperidin-1-yl)methyl)-N-(3-methoxyphenyl)pyrrolo[1,2- f] Preparation of [1,2,4]triazin-4-amine (racemic)

Compound 247C (racemic) was prepared by a method similar to that described for 146 to this 247B. Compound 247C was passed through a 1 g SCX cartridge eluting with MeOH (16 mL) followed by 2M _NH3 in MeOH (16 mL). The eluate was concentrated in vacuo and further purified by preparative HPLC to give 26 mg (yield: 42%) of 247C as a solid. It has analytical HPLC retention time = 1.51 min (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +1 = 383.

247D.5-(((3R,4R)-4-amino-3-methoxypiperidin-1-yl)methyl)-N-(3-methoxyphenyl)pyrrolo[1,2- f] Preparation of [1,2,4]triazin-4-amine (enantiomer A)

Compound 247D was obtained from compound 247C by chiral preparative HPLC separation (Chiralpak AD, 10 micron, 2×5 cm column, 220 nm, 20 mL/min, EtOH/MeOH/DEA, 50/50/0.1). Compound 247D was a solid with >99% ee and HPLC retention time = 6.5 min (Chiralpak AD, 250 x 4.6 mm, 10 micron; EtOH/MeOH/DEA: 50/50/0.1, 0.8 mL/min, 220 nM).

247E.5-(((3S,4S)-4-amino-3-methoxypiperidin-1-yl)methyl)N-(3-methoxyphenyl)pyrrolo[1,2-f Preparation of ][1,2,4]triazin-4-amine (enantiomer B)

Compound 247E was obtained from compound 247C by chiral preparative HPLC separation (Chiralpak AD, 10 micron, 2×5 cm column, 220 nm, 20 mL/min, EtOH/MeOH/DEA, 50/50/0.1). Compound 247E was a solid with >99% ee and HPLC retention time = 8.9 min (Chiralpak AD, 250 x 4.6 mm, 10 micron; EtOH/MeOH/DEA: 50/50/0.1, 0.8 mL/min, 220 nM).

Example 248

rac-5-{[(3R,4R)-4-amino-3-methoxypiperidin-1-yl]methyl}-N-(4-fluoro-3-methoxyphenyl)pyrrolo[ 2,1-f][1,2,4]triazin-4-amine

Compound 248 was prepared from 247A by a method similar to compound 247 and had an analytical HPLC retention time = 1.18 minutes (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% aqueous methanol containing 0.1% TFA over 4 minutes, 4 mL/min, monitored at 220 nm) and LC/MSM ⁺ +1 = 401.

Example 249

嗪-2(3H)-酮(4aR, 8aR)-rel-6-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazin-5-yl)methyl) -Hexahydro-1H-pyrido[3,4-b][1,4]

Azin-2(3H)-one

249A. Preparation of (3R,4R)-rel-tert-butyl 4-amino-3-hydroxypiperidine-1-carboxylate

20% Pd(OH) ₂ /C (108 mg) was added to a stirred solution of 15A (540 mg, 2.23 mmol) in 10 mL of MeOH under _N2 . The reaction flask was purged several times with _H2 and stirred under hydrogen for 18 h. The catalyst was removed by filtration through a 4 [mu]M polycarbonate membrane and rinsed with MeOH (6 x 30 mL). The filtrate was concentrated in vacuo to give 453 mg (yield: 94%) of 249A. It has analytical HPLC retention time = 0.73 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water containing 0.2% phosphoric acid over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +1 =217.

249B. Preparation of (3R,4R)-rel-tert-butyl 4-(2-chloroacetamido)-3-hydroxypiperidine-1-carboxylate

Chloroacetyl chloride (0.17 mL, 2.08 mmol) was added dropwise to a stirring of 249A (428 mg, 1.98 mmol) and NaOAc (325 mg, 3.96 mmol) in 2.5 mL of acetone and 0.8 mL of water at 0 °C under _N2 over 5 min in the mixture. The mixture was stirred at 0°C for 10 minutes and at room temperature for 25 minutes. The mixture was diluted with 160 mL of EtOAc and washed with water (2×40 mL), saturated NaHCO ₃ solution (1×30 mL) and brine (1×30 mL). The EtOAc layer was dried ( _MgSO4 ), filtered and concentrated in vacuo to give 415 mg (Yield: 72%) of 249B. It has analytical HPLC retention time = 1.96 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water containing 0.2% phosphoric acid over 4 min, 4 ml/min, monitored at 220 nm) and flow injection MS M ^- = 291.

嗪-6(7H)-羧酸酯的制备249C.(4aR,8aR)-rel-tert-butyl 2-oxo-hexahydro-1H-pyrrolo[3,4-b][1,4]

Preparation of Ozine-6(7H)-Carboxylate

60% NaH (84.0 mg, 2.10 mmol) was added to a stirred mixture of 249B (410 mg, 14.0 mmol) in 10 mL dry THF under _N2 at room temperature. The mixture was stirred at room temperature for 45 minutes and quenched with 3 mL of saturated _NH4Cl solution. The mixture was concentrated in vacuo and diluted with 40 mL saturated NaHCO ₃ solution. The aqueous solution was extracted with EtOAc (3 x 60 mL). The combined EtOAc extracts were washed with brine (1 x 30 mL). The EtOAc layer was dried (MgSO ₄ ), filtered and concentrated in vacuo to give 344 mg (Yield: 96%) of 249C. It had an analytical HPLC retention time = 2.01 min (Chromolith SpeedROD 4.6 x 50 mm, 10-90% methanol in water containing 0.2% phosphoric acid over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ = 257 .

嗪-2(3H)-酮的制备249D.(4aR,8aR)-rel-hexahydro-1H-pyrrolo[3,4-b][1,4]

Preparation of oxazin-2(3H)-one

TFA (2.00 mL, 25.9 mmol) was added to a stirred mixture of 249C (70.0 mg, 0.0.27 _mmol ) in 1 mL _CH2Cl2 at room temperature. The mixture was stirred at room temperature for 1 hour and concentrated in vacuo to give crude 249D. This material was mixed with DMA to make a 2 ml stock solution, which was used as such in the next step reaction.

Compound 249 was prepared by a method similar to compound 146 from 249D. It has analytical HPLC retention time = 1.85 min (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=409.

Example 250

嗪-6(7H)-基)甲基)-N-(3-甲氧基苯基)吡咯并[1，2-f][1，2，4]三嗪-4-胺5-(((4aR,8aR)-rel-hexahydro-1H-pyrido[3,4-b][1,4]

Azin-6(7H)-yl)methyl)-N-(3-methoxyphenyl)pyrrolo[1,2-f][1,2,4]triazin-4-amine

1M _LiAlH4 /ether solution (0.70 mL, 0.70 mmol) was added dropwise to a stirred mixture of compound 249 (100 mg, 0.24 mmol) in 4 mL of anhydrous THF at room temperature. The mixture was stirred at room temperature for 70 minutes and quenched by adding 200 mg of Celite and 200 mg of anhydrous sodium sulfate. The mixture was stirred at room temperature for 50 minutes. Insolubles were filtered and rinsed with MeOH (3 x 15 mL). The filtrate was concentrated in vacuo and purified by preparative HPLC to obtain 78 mg (yield: 81%) of compound 250. It has analytical HPLC retention time = 1.68 min (Phenomenox S5C18-HC 4.6 x 50 mm column, 10-90% aqueous methanol containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +1=395.

Example 251

嗪-6(7H)-基)甲基)吡咯并[1，2-f][1，2，4]三嗪-4-胺N-(4-fluoro-3-methoxyphenyl)-5-(((4aR,8aR)-rel-hexahydro-1H-pyrido[3,4-b][1,4]

Azin-6(7H)-yl)methyl)pyrrolo[1,2-f][1,2,4]triazin-4-amine

Compound 251 was prepared by a method similar to compound 250 from 249D. It has analytical HPLC retention time = 1.64 min (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% methanol in water, containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=413.

Example 252

(3R, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }Methyl)-N-(methylsulfonyl)piperidine-3-carboxamide

252A. Preparation of 1-tert-butyl 3-ethyl 4-oxopiperidine-1,3-dicarboxylate

A solution of ethyl 4-oxopiperidine-3-carboxylate (16.2 g, 95 mmol) in _CHCl3 (160 mL) was treated with a solution of _NaHCO3 (9.6 g, 114 mmol) in water (170 mL). This biphasic reaction was treated with a solution of _Boc2O (20.7 g, 95 mmol) in _CHCl3 (60 mL). The resulting reaction was heated to reflux for 18 hours, cooled to room temperature, and the layers were separated. The aqueous layer was extracted with _CHCl3 (2 x 100 mL). The combined organic layers were dried (Na ₂ SO ₄ ), filtered and concentrated to give 22 g (yield: 86%) of compound 252A as an oil.

252B. Preparation of (R)-1-tert-butyl 3-ethyl 4-(1-phenylethylamino)-5,6-dihydro-pyridine-1,3(2H)-dicarboxylate

A solution of 252A (22 g, 81 mmol) and toluene (400 mL) was treated with (R)-1-phenylethylamine (12.5 mL, 97 mmol) and p-TsOH (1.5 g). The reaction mixture was heated to reflux for 23 hours using a Dean-Stark trap. The mixture was cooled to room temperature, washed with saturated aqueous NaHCO ₃ (2×200 mL) and brine (2×200 mL). _The combined organic layers were dried ( _Na2SO4 ), filtered and concentrated. The crude material was filtered through a pad of silica (100% CH ₂ Cl ₂ ) and concentrated to give 14.7 g (yield: 49%) of compound 252B as an oil.

252C. Preparation of (3S,4R)-1-tert-butyl 3-ethyl 4-((R)-1-phenylethylamino)-piperidine-1,3-dicarboxylate

A 1 L 3-neck round bottom flask equipped with a mechanical stirrer and addition funnel was charged with 252B (14.7 g, 39 mmol) along with acetonitrile (200 mL) and acetic acid (100 mL). The solution was cooled to 0 °C and Na(OAc) _3BH (33.3 g, 157 mmol) was added in three portions over two hours. The reaction was stirred at this temperature for two hours after the last addition. The mixture was then cooled to -10°C and slowly quenched by the addition of 1N NaOH (100 mL), 4N NaOH (100 mL), 6N NaOH (100 mL) followed by 50% NaOH (50 mL). The reaction was allowed to warm to room temperature and the layers were separated. The aqueous layer was extracted with _CH2Cl2 (2 x 250 mL) and the combined organic layers _{were dried} ( _Na2SO4 ), filtered and concentrated in vacuo. Purification by flash chromatography on silica gel (10% to 30% EtOAc/hexanes gradient) afforded 6.0 g (yield: 41%) of compound 252C.

252D. Preparation of (3R,4R)-1-tert-butyl 3-ethyl 4-((R)-1-phenylethylamino)-piperidine-1,3-dicarboxylate

A solution of 252C (2.90 g, 7.71 mmol) in ethanol (70 mL) was treated with 21% NaOEt in ethanol (7.5 mL). The reaction mixture was heated at 50°C for three hours, cooled to room temperature, and concentrated. The resulting oil was dissolved in dichloromethane (150 mL) and washed with 20% _NH4Cl (2 x 50 mL). The organic layer was dried ( _Na2SO4 ) and concentrated to _an oil. Purification by flash chromatography on silica gel (10% to 25% EtOAc/hexanes gradient) afforded 1.20 g (yield: 41%) of compound 252D as an oil.

252E. Preparation of (3R,4R)-1-tert-butyl 3-ethyl 4-aminopiperidine-1,3-dicarboxylate.

A solution of 252D (1.18 g, 3.13 mmol) in MeOH (31 mL) was treated with ammonium formate (1.58 g, 25.1 mmol) and 10% Pd/C. The reaction mixture was heated to reflux for 14 hours and then cooled to room temperature. The resulting solid was removed by filtration and washed with MeOH. The filtrate was dried (Na ₂ SO ₄ ) and concentrated under reduced pressure to give 0.82 g (yield: 96%) of compound 252E as an oil.

252F. Preparation of (3R,4R)-1-tert-butyl 3-ethyl 4-aminopiperidine-1,3-dicarboxylate

A solution of 252E (0.80 g, 2.94 mmol) in dichloromethane (29 mL) was treated with diisopropylethylamine (0.41 g, 3.23 mmol) at 0°C. A solution of allyl chloroformate (0.46 g, 3.83 mmol) in dichloromethane (29 mL) was added slowly over 30 minutes. The reaction mixture was stirred at 0°C for 16 hours, then slowly warmed to room temperature. The resulting solution was diluted with dichloromethane. and washed with saturated NaHCO ₃ (2×50 mL). The organic layer was dried (Na ₂ SO ₄ ) and concentrated under reduced pressure. The resulting oil was purified by flash chromatography on silica gel (20 to 25% EtOAc/hexanes) to afford 0.88 g (yield: 84%) of compound 252F as an oil.

252G. Preparation of (3R,4R)-methyl 4-(allyloxycarbonyl)piperidine-3-carboxylate

A solution of 252F (0.87 g, 2.44 mmol) in dichloromethane (12 mL) was treated with TFA (2.4 mL) at 0°C. The reaction mixture was stirred at 0°C for one hour and then slowly warmed to room temperature. Stirring was continued at this temperature for five hours, concentrated and azeotroped with MeOH and toluene. The resulting oil was purified by flash chromatography on silica gel (0 to 2% MeOH/ _CH2Cl2 ) to afford 0.47 g ( _yield : 79%) of compound 252G as an oil.

252H.(3R, 4R)-methyl 4-(allyloxycarbonyl)-1-((4-(3-methoxyphenyl-amino)pyrrolo[1,2-f][1,2 , 4] Preparation of triazin-5-yl) methyl) piperidine-3-carboxylate

252G (0.21g, 0.87mmol), 4-(3-methoxyphenylamino)-pyrrolo[1,2,4]triazin-5-ylmethyltriethylammonium bromide (0.40g, 0.87 mmol) and diisopropylethylamine (0.11 g, 0.87 mmol) in MeCN (15 mL) was heated at 60° C. for one hour then concentrated in vacuo. The resulting oil was purified by flash chromatography on silica gel (2 to 5% MeOH/ _CH2Cl2 ) to afford 0.33 g (yield: 77%) of compound 252H as _a solid.

252I.(3R, 4R)-4-(allyloxycarbonyl)-1-((4-(3-methoxyphenylamino)-pyrrolo[1,2-f][1,2,4 ] Triazin-5-yl) methyl) preparation of piperidine-3-carboxylic acid

A solution of 252H (0.33 g, 0.67 mmol) in MeOH/THF/water (3/3/1/ml) was treated with LiOH monohydrate (0.25 g, 6.7 mmol). The reaction mixture was stirred for 14 h, neutralized to pH = 7 with saturated aqueous NaHCO ₃ , then concentrated to a volume of 1 mL. The resulting slurry was dissolved in MeOH and purified by preparative HPLC (YMC ODS-A 5 μm, 20×100 mm, Solvent A 10% MeOH-90% H ₂ O-0.1% TFA, Solvent B 90% MeOH-10 % _H2O -0.1%TFA, gradient 0-100%B over 12 minutes). The desired fractions were combined and concentrated under reduced pressure to remove most of the MeOH, neutralized to pH = 7 with saturated NaHCO ₃ and extracted with EtOAc (2 x 50 mL). The combined organic layers were dried (Na ₂ SO ₄ ) and concentrated to give 0.30 g (yield: 94%) of compound 252I as a solid.

252J.(3R,4R)-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]-triazin-5-yl)methyl Preparation of )-3-(methylsulfonylcarbamoyl)piperidin-4-ylcarbamate allyl ester

A solution of 252I (0.30g, 0.63mmol) in MeCN (6.2ml) was dissolved in dimethylaminopiperidine (77mg, 0.63mmol), DECI (0.18g, 0.94mmol), followed by methanesulfonamide (0.18g, 1.88mmol )deal with. The reaction mixture was stirred for two hours, quenched with water, and concentrated. The resulting slurry was dissolved in MeOH and analyzed by preparative HPLC (YMC ODS-A 5 μm, 20×100 mm, Solvent A 10% MeOH-90% H ₂ O-0.1% TFA, Solvent B 90% MeOH-10% _H2O -0.1% TFA, gradient 0-100% B, 12 min) purification. The desired fractions were combined and concentrated in vacuo, neutralized to pH=10 with saturated aqueous NaHCO ₃ , and extracted with EtOAc (2×50 mL). The combined organic layers were dried (Na ₂ SO ₄ ) and concentrated to give 0.22 g (yield: 62%) of compound 252J as a solid.

A solution of 252J (100 mg, 0.18 mmol) in THF (4 mL, degassed with argon) was treated with Pd( _PPh3 ) ₄ (21 mg, 0.018 mmol) and _Et2NH (33 mg, 0.45 mmol). The reaction mixture was stirred for 90 minutes then concentrated. The resulting solid was dissolved in MeOH and analyzed by preparative HPLC (YMCODS-A 5 μm, 20 x 100 mm, solvent A 10% MeOH-90% _H2O -0.1% TFA, solvent B 90% MeOH-10% _H2O - 0.1% TFA, gradient 0-100% B, 12 minutes) purification. The desired fractions were concentrated, neutralized to pH = 10 with saturated aqueous NaHCO ₃ , and extracted with EtOAc (2×50 mL). The combined organic layers were dried (Na ₂ SO ₄ ) and concentrated to give 36 mg (yield: 42%) of compound 252 as a solid. It has analytical HPLC retention time = 1.62 min (Phenomenex Su C18 4.6 x 50 mm column 10-90% methanol in water containing 0.2% H ₃ PO ₄ , 4 min gradient, monitored at 220 nm), [M+H] ⁺ = 474 .

Example 253

(3R,4R)-4-amino-1-({4-[(3-ethynylphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl} Methyl)-N-methylpiperidine-3-carboxamide

253A.(3R, 4R)-methyl-4-(allyloxycarbonyl)-1-((4-(3-ethynylphenyl-amino)pyrrolo[1,2-f][1,2 , 4] Preparation of triazin-5-yl) methyl) piperidine-3-carboxylate

252G (0.24g, 1.0mmol), 4-(3-ethynylphenylamino)-pyrrolo[1,2,4]triazin-5-ylmethyltriethylammonium bromide (0.43g, 1.0 mmol) and diisopropylethylamine (0.13 g, 1.0 mmol) in MeCN (15 mL) was heated at 60°C for one hour, then concentrated in vacuo. The residue was purified by silica gel flash chromatography (2 to 5% MeOH/ _CH2Cl2 ) to obtain 0.24 g (yield: 50%) of compound 253A as a _solid .

253B.(3R,4R)-4-(allyloxycarbonyl)-1-((4-(3-ethynylphenylamino)-pyrrolo[1,2-f][1,2,4] Preparation of triazin-5-yl)methyl)piperidine-3-carboxylic acid

A solution of 253A (0.24 g, 0.50 mmol) in MeOH/THF/water (3/3/1/ml) was treated with LiOH monohydrate (0.21 g, 5.0 mmol) at room temperature. The reaction mixture was stirred for 14 h, neutralized to pH=7 with saturated aqueous NaHCO ₃ , then concentrated. The resulting slurry was dissolved in MeOH and analyzed by preparative HPLC (YMC ODS-A 5 μm, 20×100 mm, solvent A 10% MeOH-90% H ₂ O-0.1% TFA, solvent B 90% MeOH-10% H ₂ O-0.1% TFA, gradient 0-100% B, 12 minutes) purification. Desired fractions were concentrated, neutralized to pH=7 with saturated NaHCO ₃ , and extracted with EtOAc (2×50 mL). The combined organic layers were dried (Na ₂ SO ₄ ) and concentrated in vacuo to afford 0.22 g (yield: 93%) of compound 253B as a solid.

253C.(3R,4R)-1-((4-(3-ethynylphenylamino)pyrrolo[1,2-f][1,2,4]-triazin-5-yl)methyl) Preparation of allyl-3-(methylcarbamoyl)piperidin-4-ylcarbamate

六氟磷酸盐(后文称为″Bop试剂″)(0.36g，0.69mmol)和在THF中的2N甲胺(0.70毫升，1.38mmol)处理。反应混合物搅拌两小时，以水停止反应，并浓缩。所得浆状物溶解在MeOH中，并通过制备型HPLC(YMC ODS-A 5微米，20×100毫米，溶剂A 10％MeOH-90％H₂O-0.1％TFA，溶剂B 90％MeOH-10％H2O-0.1％TFA，梯度0-100％B，12分钟)纯化。浓缩所需的级分，以饱和NaHCO₃水溶液中和到pH＝10，并以EtOAc(2×50毫升)萃取。干燥(Na₂SO₄)合并的有机层并在真空中浓缩以得到0.21克(产率：92％)固体的化合物253C。A solution of 253B (0.22g, 0.46mmol) in MeCN (4.6mL) was dissolved in diisopropylethylamine (59mg, 0.46mmol), benzotriazol-1-yloxy-tris(dimethylamino)

Hexafluorophosphate (hereinafter "Bop's reagent") (0.36 g, 0.69 mmol) was treated with 2N methylamine in THF (0.70 mL, 1.38 mmol). The reaction mixture was stirred for two hours, quenched with water, and concentrated. The resulting slurry was dissolved in MeOH and analyzed by preparative HPLC (YMC ODS-A 5 μm, 20×100 mm, solvent A 10% MeOH-90% H ₂ O-0.1% TFA, solvent B 90% MeOH-10 % H2O-0.1% TFA, gradient 0-100% B, 12 minutes) purification. The desired fractions were concentrated, neutralized to pH=10 with saturated aqueous NaHCO ₃ , and extracted with EtOAc (2×50 mL). The combined organic layers were dried (Na ₂ SO ₄ ) and concentrated in vacuo to afford 0.21 g (yield: 92%) of compound 253C as a solid.

A solution of 253C (100 mg, 0.20 mmol) in THF (5 mL, degassed with argon) was treated with Pd( _PPh3 ) ₄ (23 mg, 0.020 mmol) and _Et2NH (37 mg, 0.51 mmol). The reaction mixture was stirred for 90 minutes, then concentrated. The resulting solid was dissolved in MeOH and analyzed by preparative HPLC (YMCODS-A 5 μm, 20×100 mm, solvent A 10% MeOH-90% H ₂ O-0.1% TFA, solvent B 90% MeOH-10% H ₂ O - 0.1% TFA, gradient 0-100% B, 12 minutes) purification. The desired fractions were concentrated, neutralized to pH=10 with saturated aqueous NaHCO ₃ , and extracted with EtOAc (2×50 mL). The combined organic layers were dried (Na ₂ SO ₄ ) and concentrated in vacuo to give 36 mg (yield: 42%) of compound 253 as a solid. It had analytical HPLC retention time = 1.96 min (Phenomenex Su C18 4.6 x 50 mm column 10-90% methanol in water containing 0.2% H ₃ PO ₄ , 4 min gradient, monitored at 220 nm), [M+H] ⁺ = 404.

Example 254

(3R, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }Methyl)-N-methylpiperidine-3-carboxamide

254A. Preparation of (3R,4R)-1-tert-butyl 3-ethyl 4-(benzyloxycarbonyl)piperidine-1,3-dicarboxylate

A solution of 252E (180 mg, 0.66 mmol) in _CH2Cl2 (5 mL) was treated with benzyloxychloroformate (0.1 mL, 0.73 mmol) and triethylamine (0.12 mL, 0.86 _mmol ). The reaction was stirred at room temperature for 18 _h , diluted with _CH2Cl2 (10 mL), and washed with water (2 x 10 mL), 0.1N HCl (2 x 10 mL), saturated aqueous _NaHCO3 (2 x 10 mL) Wash with brine (1 x 10 mL). The organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated to give 243 mg (yield: 91%) of compound 254A as an oil.

254B. Preparation of (3R,4R)-ethyl 4-(benzyloxycarbonyl)piperidine-3-carboxylate

A solution of 254A (243 mg, 0.60 mmol) in _CH2Cl2 (3 mL) was treated with trifluoroacetic acid ( _0.3 mL) at 0 °C. The reaction was stirred at room temperature for one hour, then concentrated to an oil. The crude amine was dissolved in EtOAc (10 mL), washed with saturated aqueous NaHCO ₃ and dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The product was purified by flash silica gel chromatography (10% MeOH/CH ₂ Cl ₂ ) to obtain 95 mg (yield: 52%) of compound 254B.

254C.(3R, 4R)-methyl 4-(benzyloxycarbonyl)-1-((4-(3-methoxyphenyl-amino)pyrrolo[1,2-f][1,2 , 4] Triazin-5-yl) methyl) piperidine-3-carboxylate

N,N-diethyl-N-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazin-5-yl)methyl) A suspension of ethanaminium bromide (1.7 g, 3.62 mmol) and 254B (1.06 g, 3.6 mmol) in acetonitrile (75 mL) was treated with DIEA (0.63 mL, 3.6 mmol) and warmed to 55 °C After 12 hours. The reaction was concentrated, dissolved in EtOAc (100 mL), and washed with water (2 x 100 mL). The crude material was dried (Na ₂ SO ₄ ), filtered and concentrated to give 1.7 g (yield: 89%) of compound 254C.

254D.(3R, 4R)-4-(Benzyloxycarbonyl)-1-((4-(3-methoxyphenylamino)-pyrrolo[1,2-f][1,2,4 ] Triazin-5-yl) methyl) preparation of piperidine-3-carboxylic acid

254D (1.67 g, Yield: 100%) was prepared from 254C (1.7 g, 3.13 mmol) by a similar method used for compound 252I.

254E.(3R,4R)-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazin-5-yl)methyl) Preparation of benzyl-3-(methylcarbamoyl)piperidin-4-ylcarbamate

Compound 254E (790 mg, yield: 54%) was prepared from 254D (1.44 g, 2.71 mmol) by a similar method used for compound 253C.

A solution of 254E (1.66 g, 3.13 mmol) in MeOH (50 mL) was purged with argon for 30 min. 5% Pd/C (300 mg) was added. The reaction mixture was stirred under hydrogen for three hours, then filtered through a pad of Celite. The filtrate was concentrated in vacuo to afford 1.21 g (yield: 94%) of compound 254 as a solid. It has analytical HPLC retention time = 1.57 min (YMC S5 ODS 4.6 x 50 mm, 10-90% methanol in water containing 0.2% _H3PO4 , ₄ min gradient, monitored at 220 nm), [M+H] ⁺ = 410 .

Example 255

(3R, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidine-3-carboxylic acid

Compound 255 (33 mg, yield: 89%) was prepared from 254D (50 mg, 0.094 mmol) by a similar method used for compound 254. It had an analytical HPLC retention time = 1.54 min (YMCS5 ODS 4.6 x 50 mm, 10-90% methanol in water containing 0.2% _H3PO4 , ₄ min gradient, monitored at 220 nm). [M+H] ⁺ =397.

Example 256

(3R, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }methyl)piperidine-3-carboxamide

256A.(3R, 4R)-3-((3,4-dimethoxybenzyl)carbamoyl)-1-((4-(3-methoxyphenylamino)pyrrolo[1,2 Preparation of -f][1,2,4]triazin-5-yl)methyl)piperidin-4-ylcarbamate benzyl ester

A solution of 254D (150 mg, 0.23 mmol) in DMF (5 ml) was prepared with 3,4-dimethoxybenzylamine (77 mg, 0.46 mmol), DIEA (80 microliters, 0.46 mmol) and Bop reagent (132 mg, 0.25mmol) treatment. The reaction was stirred at room temperature for four hours, then poured into EtOAc (25 mL). The mixture was washed with saturated aqueous NaHCO ₃ (3×25 mL), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (3% MeOH/CH ₂ Cl ₂ ) to afford 174 mg (yield: 95%) of compound 256A.

A solution of 256A (50 mg, 0.06 mmol) in TFA (3 mL) was stirred at room temperature for 5 days. The reaction was concentrated and purified by preparative HPLC to afford 7 mg (yield: 30%) of compound 256 as a solid. It has analytical HPLC retention time = 1.62 min (YMC S5 ODS 4.6 x 50 mm, 10-90% methanol in water containing 0.2% _H3PO4 , ₄ min gradient, monitored at 220 nm), [M+H] ⁺ = 396 .

Example 257

(3S, 4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }Methyl)-N-(methylsulfonyl)piperidine-3-carboxamide

257A. Preparation of (3S,4R)-1-tert-butyl 3-methyl 4-aminopiperidine-1,3-dicarboxylate

Compound 257A (447 mg, 63%) was prepared by a similar method used for 252E.

257B. Preparation of (3S,4R)-1-tert-butyl 3-methyl 4-(benzyloxycarbonyl)piperidine-1,3-dicarboxylate

A solution of 257A (447 mg, 1.7 mmol) in _CH2Cl2 (20 _mL ) was treated with triethylamine (0.3 mL, 2.2 mmol) followed by benzyl chloroformate (0.27 mL, 1.9 mmol) at room temperature. The reaction mixture was stirred for 18 hours, then washed with water (25 mL). The aqueous layer was extracted with _CH2Cl2 (25 mL) and the combined organics were washed with _saturated aqueous _NaHCO3 , 0.1N HCl and brine. The organic layer was dried ( _Na2SO4 ), filtered and concentrated in vacuo to _an oil. The crude material was purified by flash chromatography on silica gel (30% EtOAc/hexanes) to give 411 mg (yield: 75%) of compound 257B as an oil.

257C. Preparation of (3S,4R)-methyl 4-(benzyloxycarbonyl)piperidine-3-carboxylate

A solution of 257B (411 mg, 1.04 mmol) in _CH2Cl2 (5 mL) was treated with TFA ₍ 0.5 mL) at room temperature. The reaction mixture was stirred for 5.0 hours, then concentrated in vacuo. The residue was dissolved in EtOAc (10 mL) and washed with saturated aqueous NaHCO ₃ . The organics were dried ( _Na2SO4 ), filtered and concentrated in vacuo to give 365 mg of 257C as _a solid.

257D.(3S, 4R)-methyl 4-(benzyloxycarbonyl)-1-((4-(3-methoxyphenyl-amino)pyrrolo[1,2-f][1,2 , 4] Preparation of triazin-5-yl) methyl) piperidine-3-carboxylate

257C (292mg, 0.62mmol) and N,N-diethyl-N-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazine A suspension of -5-yl)methyl)ethylammonium bromide (292 mg, 0.62 mmol) in acetonitrile (10 mL) was treated with DIEA (0.2 mL, 1.24 mmol) at room temperature. The reaction mixture was warmed to 55°C for 3.0 hours, then concentrated to dryness in vacuo. The crude residue was purified by flash chromatography on silica gel (30% EtOAc/hexanes) to afford 269 mg (yield: 80%) of 257D.

257E.(3S,4R)-4-(Benzyloxycarbonyl)-1-((4-(3-methoxyphenylamino)-pyrrolo[1,2-f][1,2,4 ] Triazin-5-yl) methyl) preparation of piperidine-3-carboxylic acid

A solution of 257D (200 mg, 0.37 mmol) in THF/MeOH (1:1, 4 mL) was treated with LiOH monohydrate (30 mg, 0.74 mmol) in water (1 mL) at room temperature. The reaction mixture was stirred for eight hours, then concentrated to 1 mL. The residue was diluted with water (10 mL) and extracted with EtOAc (3 x 10 mL). The organics were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give 200 mg (yield: 100%) of compound 257E as a solid.

257F.(3S, 4R)-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazin-5-yl)methyl) Preparation of -3-(methylsulfonylcarbamoyl)piperidin-4-ylcarbamate acid (carbamate acid)

A solution of 257E (30 mg, 0.06 mmol) in DMF (2 mL) was treated with methanesulfonamide (11 mg, 0.11 mmol), DMAP (7 mg, 0.06 mmol) and EDAC (13 mg, 0.07 mmol). The reaction mixture was stirred at room temperature for 48 hours. The resulting suspension was diluted with EtOAc (10 mL), washed with brine (3 x 10 mL), saturated aqueous NaHCO ₃ (2 x 10 mL), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give 35 mg 257E, which was used without further purification.

A solution of 257F (35 mg) in MeOH (3 mL) was treated with 10% Pd/C (15 mg) and stirred at room temperature under hydrogen for three hours. Filter the slurry through a nylon filter and concentrate the filtrate. The crude material was purified by preparative HPLC to afford 12 mg of compound 257 as a solid. It has analytical HPLC retention time = 1.77 min (YMC S5 ODS 4.6 x 50 mm, 10-90% methanol in water containing 0.2% _H3PO4 , ₄ min gradient, monitored at 220 nm), [M+H] ⁺ = 474 .

Example 258

(3R, 4S)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl }Methyl)-N-(methylsulfonyl)piperidine-3-carboxamide

258A. Preparation of (3R,4S)-1-tert-butyl 3-methyl 4-((S)-1-phenylethylamino)-piperidine-1,3-dicarboxylate

Compound 258A was prepared by the same method as 252C using appropriate starting materials.

258B. Preparation of (3R,4S)-1-tert-butyl 3-methyl 4-aminopiperidine-1,3-dicarboxylate

Compound 258B was prepared according to the procedure described in Example 257 using appropriate starting materials.

Compound 258 was prepared from 258B by the same method as described for 257. Compound 258 has an analytical HPLC retention time = 1.77 minutes (YMC S5 ODS 4.6×50 mm, 10-90% aqueous methanol containing 0.2% H ₃ PO ₄ , 4-minute gradient, monitored at 220 nm), [M+H] ⁺ = 474.

Example 259

(3S, 4R)-4-amino-1-({4-[(3-ethynylphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl} Methyl)-N-methylpiperidine-3-carboxamide

259A. Preparation of (3S,4R)-1-tert-butyl 3-ethyl 4-aminopiperidine-1,3-dicarboxylate

A solution of 252C (2.6 g, 6.9 mmol) in EtOH (100 mL) was treated with ammonium formate (3.5 g, 55.3 mmol) and 10% Pd/C (390 mg). The reaction mixture was heated to reflux for three hours under a nitrogen atmosphere. The resulting suspension was filtered through a pad of Celite and concentrated in vacuo to afford 1.8 g (yield: 96%) of compound 259A as a solid.

259B. Preparation of ((3S,4R)-1-tert-butyl 3-ethyl 4-(allyloxycarbonyl)piperidine-1,3-dicarboxylate

A solution of 259A (900 mg, 3.3 mmol) in _CH2Cl2 (20 mL) was treated with triethylamine (0.64 mL, 4.62 mmol) and allyl chloroformate ( _0.35 mL, 3.96 mmol) at room temperature. The mixture was stirred for four hours, then washed with 0.1N HCl (2 x 10 mL), 1 N NaOH (2 x 10 mL) and brine (10 mL). The organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give 680 mg (yield: 58%) of compound 259B as an oil.

259C. Preparation of (3S,4R)-ethyl 4-(allyloxycarbonyl)piperidine-3-carboxylate

A solution of 259B (680 mg, 1.9 mmol) in _CH2Cl2 (10 mL) was treated with TFA ₍ 1 mL) at room temperature. The reaction mixture was stirred for 16 hours, then concentrated. The residue was dissolved in EtOAc (20 mL) and washed with sat. NaHCO ₃ (2×20 mL), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give 170 mg of compound 259C.

259D.(3 S, 4R)-methyl 4-(allyloxycarbonyl)-1-((4-(3-ethynylphenylamino)-pyrrolo[1,2-f][1,2 , 4] Preparation of triazin-5-yl) methyl) piperidine-3-carboxylate

259C (50mg, 0.2mmol) and N,N-diethyl-N-((4-(3-ethynylphenyl-amino)pyrrolo[1,2-f][1,2,4]tri A suspension of azin-5-yl)methyl)ethylammonium bromide (82 mg, 0.18 mmol) in acetonitrile (2 mL) was treated with DIEA (31 μL, 0.18 mmol). The mixture was heated at 65°C for 6.0 hours, cooled to room temperature and concentrated. The crude material was purified by radial chromatography (SiO ₂ , 2 mm plate, gradient 100% CH ₂ Cl ₂ to 1% MeOH/CH ₂ Cl ₂ ) to afford 63 mg (yield: 70%) of compound 259D.

259E.(3S,4R)-4-(allyloxycarbonyl)-1-((4-(3-ethynylphenylamino)-pyrrolo[1,2-f][1,2,4] Preparation of triazin-5-yl)methyl)piperidine-3-carboxylic acid

A solution of 259D (63 mg, 0.13 mmol) in THF/MeOH (1:1, 4 mL) was treated with a solution of LiOH monohydrate (17 mg, 0.39 mmol) at room temperature. The reaction mixture was stirred for 18 hours, then concentrated to a volume of 0.5 mL. The residue was diluted with water (5 mL) and brought to pH 6 with saturated aqueous _NH4Cl . The mixture was extracted with EtOAc (2 x 10 mL), the organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo to give 56 mg (yield: 92%) of compound 259E.

259F.(3S, 4R)-1-((4-(3-ethynylphenylamino)pyrrolo[1,2-f][1,2,4]triazin-5-yl)methyl)- Preparation of Allyl 3-(Methylcarbamoyl)piperidin-4-ylcarbamate

259E (56 mg, 0.12 mmol) in DMF (3 mL) was sequentially treated with methylamine (2M in THF, 0.12 mL, 0.24 mmol), DIEA (0.02 mL, 0.12 mmol) and Bop's reagent (68 mg, 0.13 mmol) solution in. The reaction mixture was stirred at room temperature for 18 hours. The resulting mixture was diluted with EtOAc (25 mL), washed with brine (3 x 15 mL), dried ( _Na2SO4 ), filtered and concentrated in _vacuo . Crude 259E (71 mg) was used without further purification.

A solution of 259F (71 mg, 0.15 mmol) in THF (3 mL) was degassed with argon and treated with diethylamine (28 mg, 0.38 mmol) and Pd( _PPh3 ) ₄ (17 mg, 0.02 mmol). The reaction was stirred under argon atmosphere for 2.0 hours, then concentrated in vacuo and purified by preparative HPLC to afford 14 mg of compound 259. It has analytical HPLC retention time = 2.10 min (YMC S5 ODS 4.6 x 50 mm, 10-90% methanol in water containing 0.2% H ₃ PO ₄ , 4 min gradient, monitored at 220 nm), [M+H] ⁺ = 404 .

Example 260

(3S, 4S)-4-amino-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazin-5-yl)methyl Base) -N-methylpiperidine-3-carboxamide

260A. Preparation of (3S,4S)-1-tert-butyl 3-methyl 4-((S)-1-phenylethylamino)-piperidine-1,3-dicarboxylate

A solution of 258A (4.50 g, 12.4 mmol) in methanol (124 mL) was treated with 25% NaOMe in methanol (8.04 mL) at room temperature. The reaction mixture was heated at 50°C for 3.0 hours, cooled to room temperature, and concentrated in vacuo. The oily residue was dissolved in dichloromethane (200 mL) and washed with 20% _NH4Cl (2 x 75 mL). The organic layer was dried over _Na2SO4 and concentrated in _vacuo . The residue was purified by flash chromatography on silica gel (10 to 25% EtOAc/hexanes) to afford 1.50 g (yield: 30%) of compound 260A as an oil.

260B. Preparation of (3S,4S)-1-tert-butyl 3-methyl 4-aminopiperidine-1,3-dicarboxylate

A solution of 260A (1.10 g, 3.03 mmol) in MeOH (31 mL) was treated with ammonium formate (1.51 g, 24.3 mmol) and 10% Pd/C (110 mg) at room temperature. The reaction mixture was heated to reflux for 14 hours, then cooled to room temperature. The solid material was removed by filtration and washed with MeOH. The filtrate was concentrated in vacuo to afford 0.75 g (yield: 96%) of compound 260B as an oil.

260C. Preparation of (3S,4S)-1-tert-butyl 3-methyl 4-(benzyloxycarbonyl)piperidine-1,3-dicarboxylate

A solution of 260B (0.75g, 2.90mmol) in dichloromethane (30ml) was treated with triethylamine (0.35g, 3.48mmol) and N-(benzyloxycarbonyloxy)succinimide ( 0.72g, 2.90mmol) treatment. The reaction mixture was allowed to warm to room temperature and stirred for 16 hours. The reaction mixture was diluted with dichloromethane, washed with 10% citric acid (2 x 50 mL), then saturated _NaHCO3 (2 x 50 mL). The organic layer was dried over Na ₂ SO ₄ and concentrated in vacuo to give 1.01 g (yield: 89%) of oily compound 260C. It was used in the next step without further purification.

260D. Preparation of 4-(Benzyloxycarbonyl)piperidine-3-carboxylic acid (3S,4S)-methyl ester

A solution of 260C (1.01 g, 2.58 mmol) in dichloromethane (50 mL) was treated with TFA (5 mL) at 0°C. The reaction mixture was stirred at 0°C for 1.0 hour then slowly warmed to room temperature and stirred for an additional 2.0 hours. The mixture was concentrated then azeotroped with MeOH and toluene. The residue was dissolved in dichloromethane and washed with saturated _NaHCO3 (2 x 50 mL). The organic layer was dried over Na ₂ SO ₄ and concentrated in vacuo to give 0.61 g (yield: 81%) of compound 260D as oil. It was used in the next step without further purification.

260E.4-(Benzyloxycarbonyl)-1-((4-(3-methoxyphenyl-amino)pyrrolo[1,2-f][1,2,4]triazine-5- base) methyl) piperidine-3-carboxylic acid (3S, 4S)-methyl ester

260D (0.18g, 0.61mmol), 4-(3-methoxyphenylamino)-pyrrolo[1,2,4]triazin-5-ylmethyltriethylammonium bromide (0.29g, 0.61 mmol) and diisopropylethylamine (79 mg, 0.61 mmol) in MeCN (6 mL) was heated at 55°C for 12 hours and concentrated in vacuo. The residue was dissolved in dichloromethane and washed with water (2 x 50 mL). The dichloromethane portion was dried over Na ₂ SO ₄ and concentrated in vacuo to give 0.33 g (yield: 99%) of compound 260E as an oil. It was used in the next step without further purification. (M+H) ⁺ =545.

260F.(3S, 4S)-4-(Benzyloxycarbonyl)-1-((4-(3-methoxyphenylamino)-pyrrolo[1,2-f][1,2,4 ] Triazin-5-yl) methyl) preparation of piperidine-3-carboxylic acid

A solution of 260E (95 mg, 0.18 mmol) in MeOH/THF/water (1/1/0.5 mL) was treated with LiOH monohydrate (75 mg, 1.8 mmol) at room temperature. The reaction mixture was stirred for 18 h, quenched with saturated _NH4Cl (5 mL), and extracted with EtOAc (3 x 15 mL). The EtOAc layer was dried over Na ₂ SO ₄ and concentrated in vacuo to give 80 mg (yield: 89%) of compound 260F as a film. It was used in the next step without further purification. Mass (M+H) ⁺ =531.

260G.(3S,4S)-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazin-5-yl)methyl) Preparation of benzyl-3-(methylcarbamoyl)piperidin-4-ylcarbamate

A solution of 260F (40 mg, 0.075 mmol) in DMF (0.8 mL) was treated with diisopropylethylamine (10 mg, 0.075 mmol), Bop's reagent (59 mg, 0.11 mmol), then 2N methylamine in THF at room temperature (0.12 mL, 0.23 mmol) was treated. The reaction mixture was stirred for 16 hours, quenched with water, and concentrated. The resulting suspension was dissolved in MeOH and analyzed by preparative HPLC (YMC ODS-A 5 μm, 20×100 mm, solvent A 10% MeOH-90% H ₂ O-0.1% TFA, solvent B 90% MeOH-10% H ₂ O-0.1% TFA, gradient 0-100% B over 12 minutes). Desired fractions were concentrated to remove most of MeOH, neutralized to pH=10 with saturated aqueous _NaHCO3 , and extracted with EtOAc (2 x 50 mL). The combined organic layers were dried over Na ₂ SO ₄ and concentrated in vacuo to give 38 mg (yield: 93%) of solid 260G. Mass (M+H) ⁺ =544.

A solution of 260G (38 mg, 0.070 mmol) in MeOH (2 mL) was treated with 5% Pd/C (10 mg) at room temperature. The reaction mixture was stirred under hydrogen for 16 hours. The catalyst was removed by filtration. The filtrate was concentrated in vacuo to obtain 25 mg (yield: 87%) of compound 260 as a solid. It had an analytical HPLC retention time = 1.71 min (Phenomenex Su C18 4.6 x 50 mm column, 10-90% methanol in water containing 0.2% _H3PO4 , ₄ min gradient, monitored at 220 nm). Mass [M+H] ⁺ = 410.

Example 261

((3R, 4R)-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazin-5-yl)methyl)- 4-((R)-1-phenylethylamino)piperidin-3-yl)methanol

261A. Preparation of (3R,4R)-1-(tert-butoxycarbonyl)-4-((R)-1-phenylethylamino)-piperidine-3-carboxylic acid

A mixture of 252D (460 mg, 1.22 mmol) and NaOEt (1.25 mL, 21 wt% in EtOH) in EtOH (10 mL) was stirred at 50 °C for 3 hours, then at room temperature for about 48 hours. The reaction mixture was concentrated in vacuo, followed by the addition of water. The mixture was acidified with 1N HCl to pH 4-5, and the solid was collected by filtration, washed with water and dried to give 300 mg (yield: 71%) of 252A. It had an analytical HPLC retention time = 2.065 min (Chromolith SpeedROD column 4.5 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 mL/min, monitored at 220 nm). Mass (M+1) ⁺ =349.

261B. Preparation of (3R,4R)-1-tert-butyl 3-methyl 4-((R)-1-phenylethylamino)-piperidine-1,3-dicarboxylate

A solution of _TMSCHN2 (0.82 mL, 1.64 mmol, 2N in hexanes) was added to _a solution of 261A (280 mg, 0.80 mmol) in 6 mL of 1:1 _CH2Cl2 /MeOH. The mixture was stirred at room temperature for 30 min, then concentrated in vacuo and purified by flash chromatography on silica gel (Hexane/EtOAc: 80:20) to give compound 261B as an oil. It had an analytical HPLC retention time = 2.187 min (Chromolith SpeedROD column 4.5 x 50 mm, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 mL/min, monitored at 220 nm). Mass (M+1) ⁺ =363.

261C. Preparation of ((3R,4R)-4-((R)-1-phenylethylamino)piperidin-3-yl)methanol

_LiBH4 (15.5 mg, 0.71 mmol) was added to a solution of 261B (270 mg, 0.75 mmol). The mixture was heated to reflux for 1.0 hours. HPLC showed some residual starting material. More _LiBH4 (15.5 mg, 0.71 mmol) was added and the mixture was heated for an additional 2.0 h. After cooling to room temperature, ice water was added, and the mixture was concentrated in vacuo to remove THF. The aqueous residue was extracted with EtOAc (x3), and the combined extracts were dried ( _Na2SO4 ) and _concentrated in vacuo. The residue was dissolved _in 2 mL _CH2Cl2 and 2 mL TFA was added. The mixture was stirred at room temperature for 30 minutes. The mixture was concentrated in vacuo, then dried under high vacuum overnight to give 261C as an oil. It has analytical HPLC retention time = 0.590 min (Chromolith SpeedROD column 4.5 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 mL/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=235. This material was used directly in the next reaction step without further purification.

261D.((3R,4R)-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]-triazin-5-yl)methyl Base)-4-((R)-1-phenylethylamino)piperidin-3-yl)methanol

Compound 261D was prepared from 261C by a method similar to that used for compound 146E. It has analytical HPLC retention time = 1.761 min (Chromolith SpeedROD column 4.5 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=487.

A mixture of 261D (160 mg, 0.33 mmol), 10% Pd/C (39 mg) and ammonium formate (166 mg, 2.63 mmol) in MeOH (15 mL) was heated at reflux for 1.0 h. After cooling to room temperature, the catalyst was removed by filtration, and the filtrate was concentrated in vacuo. The residue was dissolved in water, basified with aqueous NaHCO ₃ and extracted with EtOAc (3x). The combined extracts were dried (Na ₂ SO ₄ ) and concentrated in vacuo to give 64 mg (yield: 51%) of compound 261 (64 mg, 51%) as a solid. It has analytical HPLC retention time = 1.137 min (Chromolith SpeedROD column 4.5 x 50 mm, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 mL/min, monitored at 220 nm) and LC/MSM ⁺ +1 =383.

Example 262

rac-(3R,4R)-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazin-5-yl}methyl Base) piperidine-3,4-diamine

262A. Preparation of 4-(benzyloxycarbonyl)-3-hydroxypiperidine-1-carboxylic acid (3R,4R)-tert-butyl ester

_Et3N (1.08 mL, 7.76 mmol) followed by Cbz-OSu (1.69 g, 6.80 mmol) were added to a stirred mixture of compound 249A (1.40 g, 6.47 mmol) in 10 mL _{of CH2Cl2} . The reaction mixture was stirred at room temperature for 16 hours, then diluted with 300 mL of EtOAc. The organic layer was washed with 5% citric acid solution (2 x 40 mL), ₅ % _K2CO3 solution (2 x 40 mL), brine (40 mL), and dried ( _MgSO4 ). The residue was filtered and concentrated in vacuo to obtain 2.25 g (yield: 99%) of compound 262A. It has analytical HPLC retention time = 3.02 minutes (Phenomenox S5C18-HC 4.6 x 50 mm column, 10-90% methanol in water, containing 0.1% TFA, over 4 minutes, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +1 = 351.

262B/C. Preparation of 3-azido-4-(benzyloxycarbonyl)piperidine-1-carboxylic acid (3R,4R)-tert-butyl ester

262B(cis) 262C(trans)

Diastereomer A Diastereomer B

Methanesulfonyl chloride (0.49 mL, 6.36 mmol) was added to compound 262A (2.23 g, 6.36 mmol) and _Et3N (1.20 mL, 0.83 mmol) in 30 mL CH under nitrogen at 0 °C over 5 min. ₂ in a stirred solution in Cl ₂ . The mixture was stirred at 0°C for 35 minutes and then diluted with 40 _mL of _CH2Cl2 . The mixture was washed with water (2 x 25 mL), brine (20 mL) and dried ( _MgSO4 ). The mixture was filtered and concentrated in vacuo to give crude mesylate salt. _NaN3 (1.65 g, 25.5 mmol) was added to the methanesulfonate in 20 mL of DMSO. The mixture was heated at 90°C for 17 hours and cooled to room temperature. The mixture was diluted with 200 mL EtOAc and washed with water (4 x 200 mL), saturated _NaHCO3 solution (40 mL), brine (40 mL) and dried ( _MgSO4 ). Filtration, concentration in vacuo, followed by flash chromatography on silica gel (15-50% EtOAc in hexanes) afforded 696 mg (29%) of 262B (diastereoisomer A, Rf = 0.65) and 262C (non- Enantiomer B, Rf = 0.70). 262B has analytical HPLC retention time = 3.51 minutes (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% aqueous methanol containing 0.1% TFA over 4 minutes, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +1 = 376. 262C has an analytical HPLC retention time = 3.51 minutes (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% methanol in water, containing 0.1% TFA, over 4 minutes, 4 ml/min, at 220nm monitoring) and LC/MS M ⁺ +1 = 376.

262D.(3R,4R)-3-azido-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazine-5 Preparation of -yl)methyl)piperidin-4-ylcarbamate benzyl ester

262D

Compound 262D was prepared from compound 262B by a method similar to that described for compound 247A. It has an analytical HPLC retention time = 2.96 minutes (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% aqueous methanol containing 0.1% TFA over 4 minutes, 4 ml/min, monitored at 220 nm) and LC/ MS M ⁺⁺ 1=528.

Compound 262 was prepared from compound 262D by a method similar to that described for compound 249A. It has analytical HPLC retention time = 1.27 min (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=368.

Example 263

rac-N-[(3R,4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]tri oxazin-5-yl}methyl)piperidin-3-yl]urea

263A. (3R, 4R)-3-amino-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazin-5-yl ) Preparation of methyl)piperidin-4-ylbenzylcarbamate (chiral, diastereoisomer A):

Compound 263A was prepared from compound 262C by a method similar to that described for compound 146E. It has analytical HPLC retention time = 2.71 min (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% methanol in water, containing 0.1% TFA, over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=502.

263B.(3R, 4R)-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazin-5-yl)methyl) Preparation of benzyl-3-ureidopiperidin-4-ylcarbamate

Trichloroacetylisocyanate (28.9 mg, 0.18 mmol) was added to a stirred mixture of compound 263A (77.0 mg, 0.15 mmol) in ₂ mL of _CH2Cl2 at 0 °C. The mixture was stirred at 0°C for 30 minutes, and 1 ml of methanol was added. This mixture was then concentrated in vacuo to give a crude oil. This crude material was dissolved in 3 mL _of methanol and 2 mL of 20% _K2CO3 solution was added. The mixture was stirred at room temperature for 2 hours, then diluted with 10 mL of water. It was concentrated in vacuo to remove methanol then extracted with EtOAc (3 x 15 mL). The combined EtOAc extracts were washed with brine (10 mL) and dried ( _MgSO4 ). Filtration followed by concentration in vacuo gave 70 mg (yield: 84%) of compound 263B. It has analytical HPLC retention time = 2.51 min (Phenomenox S5C18-HC 4.6 x 50 mm column, 10-90% aqueous methanol containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +1=545.

Compound 263 was prepared from compound 263B by a method similar to that described for compound 249A. It has analytical HPLC retention time = 1.32 min (Phenomenox S5°C 18-HC 4.6 x 50 mm column, 10-90% aqueous methanol containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/ MS M ⁺⁺ 1=411.

Example 264

rac-N-[(3R,4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]tri Azin-5-yl}methyl)piperidin-3-yl]methanesulfonamide

264A. Preparation of benzyl 7-(methylsulfonyl)-3,7-diaza-bicyclo[4.1.0]heptane-3-carboxylate (racemic)

Triethylamine (0.74 mL, 5.31 mmol) followed by methanesulfonyl chloride (0.18 mL, 2.30 mmol) was added to benzyl 3,7-diaza-bicyclo[4.1.0]heptane-3-carboxylate (410 mg, 1.77 mmol, prepared as described in Tetrahedron Letters, 43(23), 4289-4293, ₂₀₀₂ ) in a stirred mixture in 5 mL _CH2Cl2 . The mixture was stirred at room temperature for 2.5 hours then diluted with 120 mL of EtOAc. This mixture was washed with 5% citric acid solution (3 x 30 mL), saturated NaHCO ₃ solution (30 mL) and brine (30 mL). The organic layer was dried ( _MgSO4 ), filtered and concentrated in vacuo to give compound 264A in quantitative yield. It has analytical HPLC retention time = 2.56 minutes (Phenomenox S5C18-HC 4.6 x 50 mm column, 10-90% methanol in water, containing 0.1% TFA, over 4 minutes, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +Na=333.

264B. Preparation of (3R,4R)-rel-benzyl 4-azido-3-(methylsulfonamido)piperidine-1-carboxylate:

_NaN3 (458 mg, 7.08 mmol) was added to a stirred mixture of compound 264A (549 mg, 1.77 mmol) in 4 mL of DMSO. The mixture was stirred at room temperature for 2 hours and diluted with 80 mL of EtOAc. The mixture was washed with water (3 x 100 mL), saturated NaHCO ₃ solution (40 mL) and brine (40 mL). The EtOAc layer was dried (MgSO ₄ ), filtered and concentrated in vacuo to afford 530 mg (yield: 85%) of compound 264B. It has analytical HPLC retention time = 2.90 min (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=354.

264C. Preparation of (3R,4R)-rel-benzyl 4-amino-3-(methylsulfonamido)piperidine-1-carboxylate:

_Ph3P (900 mg, 3.43 mmol) was added to a stirred mixture of compound 264B (530 mg, 1.50 mmol) in 6 mL THF and 1 mL water. The reaction mixture was heated at 70°C for 15 hours and cooled to room temperature. The mixture was concentrated in vacuo, diluted with 15 mL of 2N HCl solution, and washed with _CHCl3 (3 x 20 mL). Basified to pH 12 by addition of 50% aqueous NaOH, saturated with NaCl, then extracted with EtOAc (3 x 25 mL). The combined EtOAc extracts were washed with brine (15 mL), dried (MgSO ₄ ), filtered and concentrated in vacuo to afford 490 mg (yield: 100%) of compound 264C. It has analytical HPLC retention time = 1.67 min (Phenomenox S5C18-HC 4.6 x 50 mm column, 10-90% aqueous methanol containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺ +1=328.

264D.(3R,4R)-rel-1-((4-(3-methoxyphenylamino)pyrrolo[1,2-f][1,2,4]triazin-5-yl)methyl Preparation of tert-butyl)-3-(methylsulfonamido)piperidin-4-ylcarbamate

_Et3N (0.63 mL, 4.50 mmol) followed by di-tert-butyl dicarbonate (390 mg, 1.80 _mmol ) were added to a stirred solution of compound 264C (490 mg, 1.50 mmol) in 6 mL _CH2Cl2 . The reaction mixture was stirred at room temperature for 3 hours. The mixture was diluted with 60 mL of EtOAc, washed with saturated NaHCO ₃ solution (2×15 mL) and brine (15 mL). The organic phase was dried ( _MgSO4 ), filtered and concentrated in vacuo to give the crude intermediate. The crude intermediate was purified by flash chromatography on silica gel (Hexane-EtOAc) to afford 131 mg of pure material. 20% Pd(OH) ₂ /C (30 mg) was added to this intermediate in 6 mL methanol under nitrogen. The reaction mixture was purged several times with hydrogen and stirred under hydrogen for 18 hours. The catalyst was removed by filtration using a 4 [mu]M polycarbonate membrane and rinsed with MeOH (4 x 10 mL). The combined filtrates were concentrated in vacuo to give 89 mg of crude amine intermediate.

Compound 264D was prepared from this intermediate by a method similar to that described for 146E. It has analytical HPLC retention time = 2.72 min (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% methanol in water containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=546.

TFA (2.5 mL, 32.4 mmol) was added to _a stirred solution of compound 264D (120 mg, 0.22 mmol) in 3 mL of _CH2Cl2 . The mixture was stirred at room temperature for 40 minutes, concentrated in vacuo, and purified by preparative HPLC to obtain 71 mg (yield: 73%) of compound 264. It has analytical HPLC retention time = 1.54 min (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% methanol in water, containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=446.

Example 265

N-[(3S,4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazine- 5-yl}methyl)piperidin-3-yl]methanesulfonamide

Preparation of compound 265A

Compound 265A was prepared from compound 262C (chiral, regioisomer B) by a method similar to that described for compound 262D. It has analytical HPLC retention time = 2.73 min (Phenomenox S5 C18-HC 4.6 x 50 mm column, 10-90% methanol in water, containing 0.1% TFA over 4 min, 4 ml/min, monitored at 220 nm) and LC/MS M ⁺⁺ 1=502.

Preparation of Compound 265B

Methanesulfonyl chloride (15.0 mg, 0.13 mmol) was added to a stirred mixture of compound 265A (60.0 mg, 0.12 mmol) and _Et3N (0.05 mL, 0.36 mmol) in 4 mL of DCM. The reaction mixture was stirred at room temperature for 20 hours and then diluted with 100 mL of EtOAc. The mixture was washed with saturated NaHCO ₃ solution (20 mL) and brine (20 mL). The EtOAc layer was dried ( _MgSO4 ), filtered and concentrated in vacuo to give compound 265B in quantitative yield of 70 mg. Compound 265B has an analytical HPLC retention time = 2.61 minutes (Phenomenox S5 C18-HC 4.6 × 50 mm column, 10-90% methanol in water, containing 0.1% TFA, over 4 minutes, 4 ml/min, monitored at 220 nm) and LC/ MS M ⁺⁺ 1=580.

Compound 265 was prepared from compound 265B (chiral, regioisomer B) by a method similar to that described for compound 249A. The structure of Compound 265 was determined by comparing ^{its 1} H-NMR with that of Compound 264. This compound has an analytical HPLC retention time = 1.50 minutes (Phenomenox S5 C18-HC 4.6 × 50 mm column, 10-90% methanol in water, containing 0.1% TFA, over 4 minutes, 4 ml/min, monitored at 220 nm) and LC/ MS M ⁺⁺ 1=446.

Example 266

N-[(3R,4R)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazine- 5-yl}methyl)piperidin-3-yl]methanesulfonamide (enantiomer A)

266 (enantiomer A)

Compound 266 was obtained from 264 by chiral preparative HPLC separation as the first eluting peak with ee > 99%. Compound 127A had HPLC retention time = 6.3 min (Chiral Pak, AD250 x 4.6 mm column, 10 micron, 220 nM, 0.8 mL/min, EtOH as eluent). LC/MSM ⁺ +1 = 446.

Example 267

N-(3S,4S)-4-amino-1-({4-[(3-methoxyphenyl)amino]pyrrolo[2,1-f][1,2,4]triazine-5 -yl}methyl)piperidin-3-yl]methanesulfonamide (enantiomer B)

267 (enantiomer B)

Compound 267 was obtained from 264 by chiral preparative HPLC separation with a second eluting peak with ee > 99%. Compound 267 had HPLC retention time = 7.9 min (Chiral Pak, AD250 x 4.6 mm column, 10 microns, 220 nM, 0.8 mL/min, EtOH as eluent). LC/MSM ⁺ +1 = 446.

Claims (9)

1. the compound of following formula or its pharmacy acceptable salt,

Wherein

X is-NH-;

R ²Be aryl or substituted aryl, wherein said substituted aryl is replaced by one to four following substituting group: alkyl, alkynyl, halogen or alkoxyl group, wherein said aryl are phenyl;

R ⁶, R ^6aAnd R ^6bBe independently selected from one or more hydrogen ,-NH ₂, OH, alkoxyl group ,-CONR ⁴R ⁵,-NR ⁴SO ₂Alkyl ,-NR ⁴SO ₂NR ⁴R ⁵,-OCONR ⁴R ⁵,-NH alkyl and-the NHCO alkyl;

R ³, R ⁴And R ⁵Be independently selected from hydrogen and alkyl; With

N is 1,

Moieties in wherein said alkyl or the described alkoxyl group has 1 to 7 carbon atom, and described alkynyl has 2 to 8 carbon atoms.

2. the compound or its pharmacy acceptable salt that have following formula

3. be selected from following compound:

5-[(4-amino-piperidino) methyl]-N-(3-chloro-4-fluorophenyl) pyrrolo-[2,1-f] [1,2,4] triazine-4-amine,

5-[(4-amino-piperidino) methyl]-N-phenylpyrrole [2,1-f] [1,2,4] triazine-4-amine also,

5-[(4-amino-piperidino) methyl]-N-(3-p-methoxy-phenyl) pyrrolo-[2,1-f] [1,2,4] triazine-4-amine,

5-[(4-amino-piperidino) methyl]-N-(3-ethynyl phenyl) pyrrolo-[2,1-f] [1,2,4] triazine-4-amine,

5-[(4-amino piperidine-1-yl) methyl]-N-(4-fluoro-3-p-methoxy-phenyl) pyrrolo-[2,1-f] [1,2,4] triazine-4-amine,

(3R, 4R)-4-amino-1-[[4-[(3-chloro-4-fluorophenyl) amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl] methyl] piperidines-3-alcohol,

(3S, 4S)-4-amino-1-[[4-[(3-chloro-4-fluorophenyl) amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl] methyl] piperidines-3-alcohol,

(3R, 4R)-4-amino-1-[[4-[(3-p-methoxy-phenyl) amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl] methyl] piperidines-3-alcohol,

(3S, 4S)-4-amino-1-[[4-[(3-p-methoxy-phenyl) amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl] methyl] piperidines-3-alcohol,

(3R, 4R)-4-amino-1-[[4-[(3-methoxyl group-4-fluorophenyl) amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl] methyl] piperidines-3-alcohol,

(3R, 4R)-4-amino-1-(the 4-[(3-ethynyl phenyl)-amino] pyrrolo-[2,1-f]-[1,2,4]-triazine-5-yl }-methyl) piperidines-3-alcohol,

(3R, 4R)-4-amino-1-(the 4-[(3-ethoxyl phenenyl)-amino]-pyrrolo-[2,1-f]-[1,2,4] triazine-5-yl }-methyl) piperidines-3-alcohol

(3R, 4R)-4-amino-1-(4-[(3-methoxyl group-4-methyl-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl }-methyl) piperidines-3-alcohol,

(3R, 4R)-4-amino-1-(the 4-[(3-bromophenyl) and amino] pyrrolo-[2,1-f] [1,2,4]-triazine-5-yl } methyl)-piperidines-3-alcohol,

(3R, 4R)-4-amino-1-(4-[(3-fluoro-5-methoxyl group-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl }-methyl) piperidines-3-alcohol,

(3S, 4R)-4-amino-1-(the 4-[(3-p-methoxy-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl) piperidines-3-alcohol,

(3R, 4S)-4-amino-1-(the 4-[(3-p-methoxy-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl) piperidines-3-alcohol,

(3S, 4R)-4-amino-1-(the 4-[(3-chloro-phenyl-) amino]-pyrrolo-[2,1-f] [1,2,4]-triazine-5-yl } methyl)-piperidines-3-alcohol,

(3S, 4R)-4-amino-1-(4-[(3-chloro-4-fluoro-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl }-methyl) piperidines-3-alcohol,

(3S, 4R)-4-amino-1-(the 4-[(3-ethynyl phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl) piperidines-3-alcohol,

(3R, 4S)-4-amino-1-(the 4-[(3-ethynyl phenyl)-amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl) piperidines-3-alcohol,

(3R, 4S)-4-amino-1-(the 4-[(3-chloro-phenyl-) and amino] pyrrolo-[2,1-f] [1,2,4]-triazine-5-yl } methyl)-piperidines-3-alcohol,

Carboxylamine (3R, 4R)-4-amino-1-(the 4-[(3-p-methoxy-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl) piperidines-3-base ester,

Carboxylamine (3R, 4R)-4-amino-1-(the 4-[(3-ethynyl phenyl)-amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl }-methyl) piperidines-3-base ester,

Carboxylamine (3R, 4R)-4-amino-1-(4-[(3-chloro-4-fluoro-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl }-methyl) piperidines-3-base ester,

Carboxylamine (3S, 4R)-4-amino-1-(4-[(3-chloro-4-fluoro-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl }-methyl) piperidines-3-base ester,

Carboxylamine (3S, 4R)-4-amino-1-(the 4-[(3-ethynyl phenyl)-amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl }-methyl) piperidines-3-base ester,

(3S, 4R)-4-amino-1-(the 4-[(3-p-methoxy-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl)-3-methyl piperidine-3-alcohol,

(3R/S, 5R/S)-4-amino-1-(the 4-[(3-p-methoxy-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl) piperidines-3, the 5-glycol,

(3S, 5S)-4-amino-1-(4-[(4-fluoro-3-methoxyl group-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl }-methyl) piperidines-3, the 5-glycol,

(3R, 5R)-4-amino-1-(the 4-[(3-ethynyl phenyl)-amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl) piperidines-3, the 5-glycol,

5-{[(3R, 4R)-4-amino-3-methoxyl group piperidines-1-yl] methyl }-N-(3-p-methoxy-phenyl) pyrrolo-[2,1-f] [1,2,4] triazine-4-amine,

(3R, 4R)-4-amino-1-(the 4-[(3-p-methoxy-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl)-N-(methyl sulphonyl) piperidines-3-methane amide,

(3R, 4R)-4-amino-1-(the 4-[(3-ethynyl phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl)-N-methyl piperidine-3-methane amide,

(3R, 4R)-4-amino-1-(the 4-[(3-p-methoxy-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl)-N-methyl piperidine-3-methane amide,

(3R, 4R)-4-amino-1-(the 4-[(3-p-methoxy-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl) piperidines-3-methane amide,

((3R, 4R)-1-((4-(3-p-methoxy-phenyl amino) pyrrolo-[1,2-f] [1,2,4] triazine-5-yl) methyl)-4-((R)-1-phenylethyl amino) piperidines-3-yl) methyl alcohol,

N-[(3R, 4R)-4-amino-1-(the 4-[(3-p-methoxy-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl) piperidines-3-yl] urea,

N-[(3R, 4R)-4-amino-1-({ 4-[(3-p-methoxy-phenyl) amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl methyl) piperidines-3-yl] Toluidrin and

N-[(3S, 4R)-4-amino-1-(the 4-[(3-p-methoxy-phenyl) and amino] pyrrolo-[2,1-f] [1,2,4] triazine-5-yl } methyl) piperidines-3-yl] Toluidrin,

Or its pharmacy acceptable salt.

4. pharmaceutical composition, it comprises the compound and the pharmaceutically acceptable carrier of one or more claims 1.

5. pharmaceutical composition, it comprises the compound and the pharmaceutically acceptable carrier of claim 2.

6. pharmaceutical composition, it comprises the compound and the pharmaceutically acceptable carrier of one or more claims 3.

7. one or more are used for the treatment of purposes in the medicine of cancer according to the compound of claim 1 in preparation.

8. one or more are used for suppressing the purposes of the medicine of receptor tyrosine kinase activity in preparation according to the compound of claim 1.

9. purposes according to Claim 8, wherein said receptor tyrosine kinase is selected from HER1, HER2 and HER4.

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